Steroid Derivative FXR Agonist

ABSTRACT

The present invention relates to a compound represented by formula (I), a tautomer thereof or a pharmaceutically acceptable salt thereof, and relates to applications thereof in the preparation of drugs for treating FXR related diseases.

REFERENCE TO RELATED APPLICATIONS

The present application is a U.S. national stage of PCT/CN2017/072567filed on Jan. 25, 2017, which claims the benefit of Chinese PatentApplication No. 201610061293.3 filed on Jan. 28, 2016 and Chinese PatentApplication No. 201610331759.7 filed on May 18, 2016 in the StateIntellectual Property Office of the P. R. China.

FIELD OF THE INVENTION

The present invention relates to a compound represented by formula (I),a tautomer thereof or a pharmaceutically acceptable salt thereof, andrelates to use thereof in the preparation of drugs for treatingFXR-related diseases.

BACKGROUND OF THE INVENTION

Farnesoid X Receptor (FXR) is an orphan nuclear receptor originallyidentified from a liver cDNA library of rat (B M. Forman, et al., Cell,81: 687-693(1995)), which is most closely related to insect ecdysonereceptor. FXR is a member of the ligand-activated transcription factornuclear receptor family including receptors for steroid, retinoids, andthyroid hormones (D J. Mangelsdorf, et al., Cell, 83: 841-850(1995)).Northern and in situanalyses show that FXR is greatly expressed in theliver, intestine, kidney, and adrenal gland (BM. Forman et al., Cell,81: 687-693(1995) and W. Seol et al., Mol. Endocrinnol. 9: 72-85(1995)).FXR binds to DNA after being formed a heterodimer with 9-cis retinoicacid receptor (RXR). FXR/RXR heterodimer preferentially binds toelements composed of two nuclear receptor half sites of the consensusAG(G/T)TCA, which is organized as an inverted repeat and separated by asingle nucleotide (IR-1 motif) (BM. Forman, et al., Cell, 81:687-693(1995)). However, these compounds fail to activate mouse andhuman FXR, resulting in the nature of the endogenous FXR ligand beinguncertain. Several naturally-occurring bile acids bind to and activateFXR at physiological concentrations (PCT WO 00/37077, published on Jun.29, 2000)). As discussed therein, the bile acids that serve as FXRligands comprise chenodeoxycholic acid (CDCA), deoxycholic acid (DCA),lithocholic acid (LCA), and taurine and glycine conjugates of these bileacids. WO-2005082925 discloses the use of INT747 in the preparation ofdrugs for treating FXR-related diseases.

SUMMARY OF THE INVENTION

The object of the present invention is to provide a compound representedby formula (I),

wherein,ring A is selected from 5- to 12-membered aryl, 5- to 12-memberedheteroaryl containing 1 to 2 heteroatoms, 5- to 6-membered non-aromaticheterocyclyl containing 1 to 2 heteroatoms or 5- to6-membered cycloalkyl, and said ring A is optionally substituted with 1,2 or 3 R_(a), or with 1, 2 or 3 oxo groups, and said heteroatom isselected from N, O or S;L is selected from C₁₋₈ alkyl, C₁₋₈ heteroalkyl or C₂₋₈ alkenyl, andsaid L is optionally substituted with 1, 2 or3 R_(b), or with 1, 2 or 3 oxo groups;L₁ is selected from O, N(R_(d)), N(R_(d))S(═O)₂ or N(R_(d))S(═O);R₁ is selected from H, C₁₋₆ alkyl, C₁₋₆ heteroalkyl, 5- to 6-memberedaryl, 4- to 6-membered heteroaryl, C₃₋₆ cycloalkyl, or 3- to 6-memberedheterocycloalkyl, and said C₁₋₆ alkyl, C₁₋₆ heteroalkyl, 5- to6-membered aryl,4- to 6-membered heteroaryl, C₃₋₆ cycloalkyl, or 3- to 6-memberedheterocycloalkyl is optionally substituted with 1, 2 or 3 R_(c), or with1, 2 or 3 oxo groups;R_(a), R_(b), R_(c) and R_(d) are each independently selected from H, F,Cl, Br, I, OH, CN, NO₂, NH₂, COOH, S(═O)₂OH, C₁₋₃ alkylamino,N,N-di(C₁₋₃ alkyl)amino, C₁₋₃ alkyl, C₁₋₃ alkyloxy or C₁₋₃ alkylthio,and said C₁₋₃ alkylamino, N,N-di(C₁₋₃ alkyl)amino, C₁₋₃ alkyl, C₁₋₃alkyloxy or C₁₋₃ alkylthio is optionally substituted with 1, 2 or 3 R′;R′ is selected from F, Cl, Br, I, OH, NH₂, NO₂, CN, COOH, Me, Et, CH₂F,CHF₂, CF₃, CH₃O, CH₃S, NH(CH₃) or N(CH₃)₂;or a pharmaceutically acceptable salt thereof.

In some embodiments of the present invention, in the compoundrepresented by formula (I) or the pharmaceutically acceptable saltthereof, the above R_(a), R_(b), R_(c) and R_(d) are each independentlyselected from H, F, Cl, Br, I, COOH, S(═O)₂OH, Me, CF₃, CHF₂, CH₂F, Et,OMe, NH(CH₃) or N(CH₃)₂.

In some embodiments of the present invention, in the compoundrepresented by formula (I) or the pharmaceutically acceptable saltthereof, the above ring A is selected from phenyl, pyridyl,pyridin-2(1H)-onyl, pyrimidyl, pyrazolyl, imidazolyl, oxazolyl,thiazolyl, thienyl, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl,isoxazolyl, isothiazolyl, bicyclo[1.1.1]pentyl, benzoxazolyl,benzo[d]isoxazolyl, indazolyl, indolyl, quinolinyl, isoquinolinyl,quinazolinyl, 1H-pyrrolo[2,3-B]pyridyl, indolizinyl, benzothiazolyl orbenzothienyl, and said ring A is optionally substituted with 1, 2 or 3R_(a).

In some embodiments of the present invention, in the compoundrepresented by formula (I) or the pharmaceutically acceptable saltthereof, the above ring A is selected from

and said ring A is optionally substituted with 1, 2 or 3 R_(a).

In some embodiments of the present invention, in the compoundrepresented by formula (I) or the pharmaceutically acceptable saltthereof, the above ring A is selected from

In some embodiments of the present invention, in the compoundrepresented by formula (I) or the pharmaceutically acceptable saltthereof, the above L is selected from C₁₋₄ alkyl, C₁₋₄ alkoxy, C₁₋₄alkylthio, C₁₋₄ alkylamino, C₁₋₄ alkyl-C(═O)NH—, C₂₋₄ alkenyl or C₁₋₃alkyl-O—C₁₋₃ alkyl, and said L is optionally substituted with 1, 2 or 3R_(b).

In some embodiments of the present invention, in the compoundrepresented by formula (I) or the pharmaceutically acceptable saltthereof, the above L is selected from

and said L is optionally substituted with 1, 2 or 3 R_(b).

In some embodiments of the present invention, in the compoundrepresented by formula (I) or the pharmaceutically acceptable saltthereof, the above L is selected from

In some embodiments of the present invention, in the compoundrepresented by formula (I) or the pharmaceutically acceptable saltthereof, the above L is selected from

In some embodiments of the present invention, the above L₁ is selectedfrom O, NH, NHS(═O)₂ and NHS(═O).

In some embodiments of the present invention, in the compoundrepresented by formula (I) or the pharmaceutically acceptable saltthereof, the above R₁ is selected from H, C₁₋₃ alkyl, C₃₋₆ cycloalkyl,phenyl, pyridyl, pyridazinyl, pyrazinyl, imidazolyl, pyrazolyl,thiazolyl, isothiazolyl, oxazolyl, isoxazolyl or thienyl, and said C₁₋₃alkyl, C₃₋₆ cycloalkyl, phenyl, pyridyl, pyridazinyl, pyrazinyl,imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl orthienyl is optionally substituted with 1, 2 or 3 R_(c), and said R_(c)is as above defined.

In some embodiments of the present invention, in the compoundrepresented by formula (I) or the pharmaceutically acceptable saltthereof, the above R₁ is selected from H, Me,

As preferred embodiments, the compound represented by formula (I) or thepharmaceutically acceptable salt thereof is selected from the compoundrepresented by the following formula (II):

wherein,L₂ is selected from C₁₋₆ alkyl, C₁₋₆ heteroalkyl or C₂₋₆ alkenyl;ring B is selected from benzene ring or 5- to 6-membered heteroaromaticring containing 1 to 2 heteroatoms, and said heteroatom is selected fromN, O or S;said ring B is optionally substituted with 1, 2 or 3 R₂, or with 1 oxogroup, and said R₂ is selected from F, Cl, Br, I, OH, CN, NO₂, NH₂,COOH, S(═O)₂OH, C₁₋₃ alkylamino, N,N-di(C₁₋₃ alkyl)amino, C₁₋₃ alkyl,C₁₋₃ alkyloxy or C₁₋₃ alkylthio;L₁ is selected from O, NH, NHS(═O)₂ or NHS(═O);R₁ is selected from H, C₁₋₆ alkyl, C₁₋₆ heteroalkyl, 5- to 6-memberedaryl, 4- to 6-membered heteroaryl, C₃₋₆ cycloalkyl, or 3- to 6-memberedheterocycloalkyl, and said C₁₋₆ alkyl, C₁₋₆ heteroalkyl, 5- to6-membered aryl,4- to 6-membered heteroaryl, C₃₋₆ cycloalkyl, or 3- to 6-memberedheterocycloalkyl are optionally substituted with 1, 2, or 3 R_(c), orwith 1, 2 or 3 oxo groups, and said R_(c) is selected from F, Cl, Br, I,OH, CN, NO₂, NH₂, COOH or S(═O)₂OH;or a pharmaceutically acceptable salt thereof.

In some embodiments of the present invention, in the compoundrepresented by formula (II) or the pharmaceutically acceptable saltthereof, L₂ is selected from —(CH₂)₁₋₆—, —(CH₂)₀₋₄—X—(CH₂)₀₋₄— or—(CH₂)₀₋₄—Y—(CH₂)₀₋₄—, and said X is selected from NH, O or S, and saidY is selected from —CH═CH—, providing that the chain length of L₂ isselected from 1 to 6, preferably 2 to 4.

In some embodiments of the present invention, in the compoundrepresented by formula (II) or the pharmaceutically acceptable saltthereof, L₂ is selected from —CH₂CH₂—, —CH₂CH₂CH₂—, —CH═CH—, —CH₂CH═CH—,—CH═CHCH₂—, —CH₂CH₂O—, —CH₂CH₂CH₂O—, —CH₂CH₂NH—, —CH₂CH₂CH₂NH—,—CH₂CH₂S— or —CH₂CH₂CH₂S—, preferably from —CH₂CH₂—, —CH₂CH₂CH₂—,—CH═CH—, —CH₂CH₂O—, —CH₂CH₂CH₂O— or —CH₂CH₂NH—.

In some embodiments of the present invention, in the compoundrepresented by formula (II) or the pharmaceutically acceptable saltthereof, ring B is selected from benzene ring, 5-membered heteroaromaticring containing 1 to 2 heteroatoms selected from N, O or S, or6-membered heteroaromatic ring containing 1 to 2 heteroatoms selectedfrom N, and said ring B is optionally substituted with 1, 2, or 3 R₂ orwith 1 oxo group, and said R₂ is as above defined.

In some embodiments of the present invention, in the compoundrepresented by formula (II) or the pharmaceutically acceptable saltthereof, ring B is selected from benzene ring, pyrazolyl, imidazolyl,oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridyl, pyridazinyl,pyrimidinyl or pyrazinyl, and said ring B is optionally substituted with1, 2 or 3 of R₂ or with 1 oxo group, and said R₂ is as above defined.

In some embodiments of the present invention, in the compoundrepresented by formula (II) or the pharmaceutically acceptable saltthereof, said ring B is optionally substituted with 1 R₂, or with 1 oxogroup, and said R₂ is selected from F, Cl, Br, I, OH, CN, NO₂, NH₂, COOHor S(═O)₂OH, preferably from F, Cl or Br.

In some embodiments of the present invention, in the compoundrepresented by formula (II) or the pharmaceutically acceptable saltthereof, the R₂-substituted or oxo-substituted position is at theortho-position relative to L₂ on ring B (a position).

In some embodiments of the present invention, in the compoundrepresented by formula (II) or the pharmaceutically acceptable saltthereof, said R₁ is selected from H, C₁₋₃ alkyl, C₃₋₆ cycloalkyl,phenyl, pyridyl, pyridazinyl, pyrazinyl, imidazolyl, pyrazolyl,thiazolyl, isothiazolyl, oxazolyl, isoxazolyl or thienyl, and said C₁₋₃alkyl, C₃₋₆ cycloalkyl, phenyl, pyridyl, pyridazinyl, pyrazinyl,imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, orthienyl is optionally substituted with 1, 2, or 3 R_(c), and said R_(c)is as above defined.

In some embodiments of the present invention, in the compoundrepresented by formula (II) or the pharmaceutically acceptable saltthereof, said R₁ is selected from H, Me,

In some embodiments of the present invention, in the compoundrepresented by formula (II) or the pharmaceutically acceptable saltthereof, the structure moiety L₁-R₁ is selected from —OH, —NHSO₂CH₃,

—NHSO₂C(CH₃)₃, —NHCH₂CH₂SO₂OH, —NHCH₂COOH,

—NHSO₂CH(CH₃)₂,

—NHCH₃, —OCH₃ or

In some embodiments of the present invention, in the compoundrepresented by formula (II) or the pharmaceutically acceptable saltthereof, the structure moiety

is selected from

and said L₂ and R₂ are as above defined.

In some embodiments of the present invention, in the compoundrepresented by formula (II) or the pharmaceutically acceptable saltthereof, the structure moiety

is selected from

The present invention also provides a compound represented by formula(III):

wherein,ring A is 5- to 12-membered aryl, 5- to 12-membered heteroaryl, or 5- to6-membered non-aromatic heterocyclyl, 5- to 6-membered cycloalkyl,optionally substituted with 1, 2 or 3 of R;L is C₁₋₆ alkyl, C₁₋₆ heteroalkyl, C₂₋₆ alkenyl, optionally substitutedwith 1, 2 or 3 of R;R is F, Cl, Br, I, OH, CN, NO₂, NH₂, or R is selected from C₁₋₃alkylamino, N,N-di(C₁₋₃ alkyl)amino, C₁₋₃ alkyl, C₁₋₃ alkyloxy, C₁₋₃alkylthio, optionally substituted with 1, 2 or 3 R′;R′ is selected from F, Cl, Br, I, OH, NH₂, NO₂, CN, COOH, Me, Et, CH₂F,CHF₂, CF₃, CH₃O, CH₃S, NH(CH₃), N(CH₃)₂;said “hetero” represents heteroatom or heteroatomic group, selected from—C(═O)NH—, N, —NH—, —C(═NH)—, —S(═O)₂NH—, —S(═O)NH—, —O—, —S—, N, ═O,═S, —C(═O)O—, —C(═O)—, —C(═S)—, —S(═O)—, —S(═O)₂—, and —NHC(═O)NH—;in any of the above cases, the number of the heteroatom or heteroatomicgroups is independently selected from 1, 2 or 3;or a pharmaceutically acceptable salt thereof.

In some embodiments of the present invention, in the compoundrepresented by formula (III) or the pharmaceutically acceptable saltthereof, the above R is selected from F, Cl, Br, I, Me, CF₃, CHF₂, CH₂F,Et, OMe, NH(CH₃), N(CH₃)₂.

In some embodiments of the present invention, in the compoundrepresented by formula (III) or the pharmaceutically acceptable saltthereof, the above ring A is phenyl, pyridyl, pyridin-2(1H)-onyl,pyrimidyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, thienyl,pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, isoxazolyl,isothiazolyl, bicyclo[1.1.1]pentyl, benzoxazolyl, benzo[d]isoxazolyl,indazolyl, indolyl, quinolinyl, isoquinolinyl, quinazolinyl,1H-pyrrolo[2,3-B]pyridyl, indolizinyl, benzothiazolyl and benzothienyl,optionally substituted with 1, 2 or 3 of R.

In some embodiments of the present invention, in the compoundrepresented by formula (III) or the pharmaceutically acceptable saltthereof, the above ring A is selected from

optionally substituted with 1, 2 or 3 R.

In some embodiments of the present invention, in the compoundrepresented by formula (III) or the pharmaceutically acceptable saltthereof, the above ring A is selected from

In some embodiments of the present invention, in the compoundrepresented by formula (III) or the pharmaceutically acceptable saltthereof, the above L is selected from C₁₋₄ alkyl, C₁₋₄ alkoxy, C₁₋₄alkylthio, C₁₋₄ alkylamino, C₁₋₄ alkyl-C(═O)NH—, C₂₋₄ alkenyl,optionally substituted with 1, 2 or 3 R.

In some embodiments of the present invention, in the compoundrepresented by formula (III) or the pharmaceutically acceptable saltthereof, the above L is selected from

optionally substituted with 1, 2 or 3 R.

In some embodiments of the present invention, in the compoundrepresented by formula (III) or the pharmaceutically acceptable saltthereof, the above L is selected from

In some embodiments of the present invention, in the compoundrepresented by formula (III) or the pharmaceutically acceptable saltthereof, the above L is selected from

The compound of the present invention is selected from:

The present invention also provides a pharmaceutical compositioncomprising a therapeutically effective amount of the above compound or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier.

The present invention also provides use of the above compound, or thepharmaceutically acceptable salt thereof, or the above pharmaceuticalcomposition in the preparation of a drug for treating Farnesoid XReceptor related diseases, cholestatic liver diseases, fibroticdiseases, hypercholesterol diseases, hypertriglyceride diseases andcardiovascular diseases.

The present invention also provides use of the above compound, or thepharmaceutically acceptable salt thereof, or the above pharmaceuticalcomposition in the preparation of a drug for treating non-alcoholicfatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH),primary biliary cirrhosis (PBC), cholestatic hepatopathy, chronic liverdisease, hepatitis C infection, alcoholic liver disease, hepaticfibrosis, primary sclerosing cholangitis (PSC), gallstone, biliaryatresia, lower urinary tract symptom and benign prostatic hyperplasia(BPH), ureteral calculi, obesity, type 2 diabetes, atherosclerosis,atherosclerotic symptom, hepatic function injury resulting fromhypercholesterolemia and hyperlipidemia.

The compound of the present invention is FXR agonist. The compound ofthe invention can be used for methods of preventing or treatingdyslipidemia or diseases associated with dyslipidemia, comprisingadministering a therapeutically effective amount of the compound of thepresent invention to a patient in need thereof.

The compound of the present invention can be used for reducing totalcholesterol level, reducing LDL cholesterol level, reducing VLDLcholesterol level, increasing HDL cholesterol level, and/or reducingtriglyceride level. Reducing triglyceride level according to the presentinvention means that the triglyceride level in subjects in need thereofis reduced below the initial triglyceride level of the subjects to beprevented or treated before taking the compounds of the presentapplication. For example, the compound of the present invention canreduce hepatic triglyceride production or hepatic triglyceride secretionby reducing fat absorption. The compound of the present invention canalso reduce serum triglyceride and hepatic triglyceride.

The compound of the present invention can be used for preventing ortreating cardiovascular diseases associated with hypertriglyceridemiaand/or hypercholesterolemia in a subject (e.g., a mammal, particularlyhuman), such as but not limited to, atherosclerosis, atheroscleroticsymptom, hypercholesterolemia, hyperlipidemia, thrombosis, coronaryartery disease, stroke or hypertensive disease.

The compound of the present invention can be used for preventing ortreating liver/biliary system diseases in a subject (e.g., a mammal,particularly human), such as but not limited to, cholestatic liverdiseases, hypercholesterol diseases, hypertriglyceride diseases orfibrotic diseases, such as but not limited to, non-alcoholicsteatohepatitis (NASH), primary biliary cirrhosis (PBC), primarysclerosing cholangitis (PSC), gallstone, non-alcoholic cirrhosis,biliary atresia, cholestatic hepatopathy, chronic liver disease,hepatitis infection (type B or C), alcoholic liver disease or hepaticfibrosis.

The compound of the present invention can be used for preventing ortreating obesity in a subject (e.g., a mammal, particularly human).

The compound of the present invention can be used for preventing ortreating diabetes, or insulin resistance, glucose intolerance relateddiseases in a subject (e.g., a mammal, particularly human).

The compound of the present invention can be used for preventing ortreating lower urinary tract symptom (surrounding area) and benignprostatic hyperplasia (BPH) or ureteral calculi in a subject (e.g., amammal, particularly human).

Another aspect of the present invention provides use of the compoundrepresented by formula I, II or III, the pharmaceutically acceptablesalt thereof or the above pharmaceutical composition in the preparationof a drug for preventing or treating a disease benefiting from FXRagonism, and said disease benefiting from FXR agonism includescardiovascular diseases, liver/biliary system diseases, obesity,diabetes, lower urinary tract symptom (surrounding area) and benignprostatic hyperplasia (BPH) or ureteral calculi.

Another aspect of the present invention provides a method for preventingor treating a disease benefiting from FXR agonism, comprisingadministering a therapeutically effective amount of the compound offormula I, II or III, the pharmaceutically acceptable salt thereof, orthe above pharmaceutical composition to a patient, and said diseasebenefiting from FXR agonism includes cardiovascular diseases,liver/biliary system diseases, obesity, diabetes, lower urinary tractsymptom (surrounding area) and benign prostatic hyperplasia (BPH) orureteral calculi.

The cardiovascular diseases include cardiovascular diseases associatedwith hypertriglyceridemia and/or hypercholesterolemia. Thecardiovascular diseases further include atherosclerosis, atheroscleroticsymptom, hypercholesterolemia, hyperlipidemia, thrombosis, coronaryartery disease, stroke or hypertension. The liver/biliary systemdiseases include cholestatic liver diseases, hypercholesterol diseases,hypertriglyceride diseases or fibrotic diseases. The liver/biliarysystem diseases further include non-alcoholic steatohepatitis (NASH),primary biliary cirrhosis (PBC), primary sclerosing cholangitis (PSC),gallstone, nonalcoholic cirrhosis, biliary atresia, cholestatichepatopathy, chronic liver disease, hepatitis infection (type B or C),alcoholic liver disease or hepatic fibrosis.

Definitions and Introductions

Unless otherwise indicated, the following terms and phrases used hereinare intended to have the following meanings. A particular term or phraseshould not be considered to be indefinite or unclear in the absence of aspecific definition, but should be interpreted as its ordinary meanings.When a trade name appears herein, it is intended to refer to thecorresponding commodity or active ingredient thereof.

The term “substituted” means that any one or more of the hydrogen atomson a specific atom are substituted by a substituent, as long as thevalence state of the specific atom is normal and the substitutedcompound is stable.

The term “oxo” means that the two hydrogen atoms on a specific atom aresubstituted by ═O. When an aromatic ring is oxo-substituted, theoxo-substituted ring may remain aromatic, or lose aromaticity. It can beunderstood that the oxo-substituted group is stable. For example, whenthe benzene ring is oxo-substituted, it can be selected from

when pyridine is oxo-substituted, it can be selected from

The term “optional” or “optionally” means that the subsequentlydescribed event or situation may occur or not, and the descriptionincludes the event or situation occurs and not. For example, an ethyl is“optionally” substituted by a halogen, meaning that the ethyl may beunsubstituted (CH₂CH₃), monosubstituted (e.g., CH₂CH₂F), polysubstituted(e.g., CHFCH₂F, CH₂CHF₂, etc.) or completely substituted (CF₂CF₃). Also,for example, “optionally, said ring B is substituted by 1, 2 or 3 R₂, orby 1 oxo group” means that ring B is substituted by 1, 2 or 3 R₂ or by 1oxo group, or ring B is not substituted by R₂ or oxo group. It can beunderstood by the skilled in the art that, for any groups containing oneor more substituents, any substitutions or substitution patterns thatare unable to exist spatially and/or cannot be synthesized will not beintroduced.

The C_(m-n) herein means that the moiety has an integer number of carbonatom in a designated range. For example, “C₁₋₆” means that the group mayhave 1, 2, 3, 4, 5 or 6 carbon atoms.

Numerical range herein means each integer in a designated range. Forexample, “C₁₋₃ alkyl” means that the group may be selected from C₁, C₂and C₃ alkyl; “C₃₋₁₀ cycloalkyl” means the group may be selected fromC₃, C₄, C₅, C₆, C₇, C₈, C₉ and C₁₀ cycloalkyl; “5- to 6-memberedcycloalkyl” means that the group may be selected from 5- and 6-memberedheterocycloalkyl.

When any variable (e.g., R) appears more than once in composition orstructure of a compound, its definition in each case is independent.Thus, for example, if a group is substituted by 0-2 R, this group may beoptionally substituted by at most two R, and R in each case hasindependent options. Furthermore, the combination of substituents and/orvariants thereof is allowed only if such combination results in stablecompounds.

When part of groups can be substituted or oxo-substituted herein, thesubstitution or oxo-substitution is independent with each other,including the cases that the groups are either substituted oroxo-substituted only, or substituted and oxo-substituted simultaneously.For example, when ring A is optionally substituted by 1, 2 or 3 R_(a),or by 1, 2 or 3 oxo groups, it includes that: 1) there are nosubstituents and oxo groups on ring A, 2) ring A is only substituted by1, 2 or 3 R_(a), 3) ring A is only substituted by 1, 2 or 3 oxo groups,4) ring A is substituted by R_(a) and oxo groups simultaneously.

In some preferred embodiments of the present application, part of moietystructures may be connected to other structures at the left end, and maybe connected to other structures at the right end at the same time.

When a dashed line or a solid line represents a linkage bond, theskilled in the art can understand by reading the present applicationthat the dashed line or solid line directionally indicates the linkagestate of the moiety structure with other structures. For example, ifring A is selected from

the leftward dashed line indicates that N atom is connected to the Lgroup of the compound represented by Formula (I), and the rightwarddashed line indicates that C atom is connected to the carbonyl moiety ofthe compound represented by formula (I); also, for example, if ring A isselected from

the leftward dashed line indicates that C atom is connected to the Lgroup of the compound represented by formula (I), and the rightwarddashed line indicates that N atom is connected to the carbonyl moiety ofthe compound represented by formula (I). The leftward dashed or solidline refers to protruding to the upper left, left, or lower left fromthe moiety structure per se; and the rightward dashed or solid linerefers to protruding to the upper right, right, or lower right from themoiety structure per se.

In some preferred embodiments of the present invention, the chain lengthrefers to the number of the atoms used to form the open chain group, andthe atoms include C atom, N atom, O atom or S atom with differentvalences. For example, when L₂ is selected from —CH₂CH₂CH₂—, the chainlength is 3; when L₂ is selected from —(CH₂)₂—O—(CH₂)₂—, the chainlength is 5; when L₂ is selected from —(CH₂)₂—CH═CH—(CH₂)₂—, the chainlength is 6.

The term “halo” or “halogen” refers to fluorine, chlorine, bromine andiodine.

The term “hydroxy” refers to —OH group.

The term “cyano” refers to —CN group.

The term “thiol” refers to —SH group.

The term “amino” refers to —NH₂ group.

The term “nitro” refers to —NO₂ group.

The term “alkoxy” refers to —O-alkyl.

The term “alkylamino” refers to —NH-alkyl.

The term “dialkylamino” refers to —N(alkyl)₂.

The term “alkylsulfonyl” refers to —SO₂-alkyl.

The term “alkylthio” refers to —S-alkyl.

The term “alkyl” refers to a hydrocarbyl with the general formula ofC_(n)H_(2n+1). The alkyl may be linear or branched. For example, theterm “C₁₋₆ alkyl” refers to an alkyl containing 1 to 6 carbon atoms (forexample, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl,3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc.). Similarly, thealkyl moieties (i.e., alkyl) of alkoxy, alkylamino, dialkylamino,alkylsulfonyl and alkylthio have the same definition as above.

The term “heteroalkyl” refers to an alkyl structure containing aheteroatom. Unless otherwise indicated, the heteroalkyl is typically analkyl containing 1 to 3 heteroatoms (preferably 1 or 2 heteroatoms)independently selected from sulfur, oxygen and/or nitrogen.

The term “alkenyl” refers to a linear or branched unsaturated aliphatichydrocarbyl having at least one double bond consisting of carbon andhydrogen atoms. Non-limiting examples of alkenyl include, but are notlimited to, vinyl, 1-propenyl, 2-propenyl, 1-butenyl, isobutenyl,1,3-butadienyl, etc. The term “linear alkenyl” refers to straight chainalkenyl.

The term “aryl” refers to an all-carbon monocyclic group with aconjugated n-electron system or a fused polycyclic aromatic ring group.For example, an aryl may have 6 to 20, 6 to 14, or 6 to 12 carbon atoms.

Non-limiting examples of aryl include, but are not limited to, phenyl,naphthyl, anthryl and 1,2,3,4-tetrahydronaphthalene, etc.

The term “heteroaryl” refers to a monocyclic or fused polycyclic systemcontaining at least one cyclic atom selected from N, O, S with theremaining cyclic atoms being C, and having at least one aromatic ring.Preferred heteroaryl has a single 4- to 8-, especially 5- to 8-memberedring, or has a plurality of fused rings containing 6 to 14, especially 6to 10 cyclic atoms. Non-limiting examples of heteroaryl include, but arenot limited to, pyrrolyl, furyl, thienyl, imidazolyl, oxazolyl,pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, quinolinyl, isoquinolinyl,tetrazolyl, triazolyl, triazinyl, benzofuranyl, benzothienyl, indolyl,isoindolyl, etc.

The term “cycloalkyl” refers to a carbocyclic ring that is fullysaturated and can exist as a monocyclic ring, bridged ring orspirocyclic ring. Unless otherwise indicated, the carbocyclic ring istypically a 3- to 10-membered ring. Non-limiting examples of cycloalkylinclude, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, norbornyl (bicyclo[2.2.1]heptyl), bicyclo[2.2.2]octyl,adamantyl etc.

The term “non-aromatic heterocyclyl” refers to a fully saturated orpartially unsaturated (but not fully unsaturated heteroaromatic)non-aromatic ring that may be present as a monocyclic ring, bicyclicring or spirocyclic ring. Unless otherwise indicated, the heterocyclicring is typically a 3- to 6-membered ring containing 1 to 3 heteroatoms(preferably 1 or 2 heteroatoms) independently selected from sulfur,oxygen, and/or nitrogen. Non-limiting examples of heterocyclyl include,but are not limited to oxiranyl, tetrahydrofuranyl, dihydrofuranyl,pyrrolidinyl, N-methyl pyrrolidinyl, dihydropyrrolyl, piperidinyl,piperazinyl, pyrazolidinyl, 4H-pyranyl, morpholinyl, thiomorpholinyl,tetrahydrothienyl, etc.

As used herein, the term “pharmaceutically acceptable” refers to thosecompounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

The term “pharmaceutically acceptable salt” refers to the salt of thecompound of the present invention, which is prepared from the compoundwith specific substituents found in the present invention and arelatively nontoxic acid or base. When the compound of the presentinvention contains relatively acidic functional groups, the baseaddition salts thereof can be obtained by contacting the neutral form ofsuch compound with a sufficient amount of base in a pure solution orsuitable inert solvent. When the compound of the present inventioncontains relatively basic functional groups, the acid addition saltsthereof can be obtained by contacting the neutral form of such compoundwith a sufficient amount of acid in a pure solution or suitable inertsolvent. Pharmaceutically acceptable salts can be mentioned as metalsalts, ammonium salts, salts formed with organic bases, salts formedwith inorganic acids, salts formed with organic acids, salts formed withbasic or acidic amino acids, etc.

Preferably, the salt is contacted with a base or acid in a conventionalmanner and the parent compound is then isolated, thereby regeneratingthe neutral form of the compound. The parent form of the compounddiffers from the various salt forms thereof in certain physicalproperties, for example, different solubilities in polar solvents.

Certain compounds of the present invention may have asymmetric carbonatoms (optical centers) or double bonds. Racemates, diastereomers,geometric isomers, and individual isomers are all included within thescope of the present invention.

The graphic representations of racemic, ambiscalemic and scalemic orenantiomerically pure compounds herein are from Maehr, J. Chem. Ed.1985, 62: 114-120. Unless otherwise stated, the absolute configurationof a stereocenter is represented by solid and broken wedges. When thecompounds described herein contain olefinic double bonds or othergeometric asymmetrical centers, unless otherwise specified, they includeE Zgeometric isomers. Likewise, all tautomeric forms are included withinthe scope of the present invention.

The compound of the present invention may exist in specific geometric orstereoisomeric forms. All such compounds envisaged by the presentinvention include cis and trans isomers, (−)- and (+)-enantiomer pairs,(R)- and (5)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, andracemic mixtures and other mixtures thereof, such as enantiomers ordiastereomers enriched mixtures, all of which fall within the scope ofthe present invention. Other asymmetric carbon atoms may be present inthe substituents such as alkyl. All these isomers and their mixtures areincluded in the scope of the present invention.

The optically active (R)- and (S)-isomers as well as the D and L isomerscan be prepared by chiral synthesis or chiral reagents or otherconventional techniques. If an enantiomer of a certain compound of thepresent invention is desired, it may be prepared by asymmetricsynthesis, or by derivatization with a chiral auxiliary, wherein theresulting diastereomeric mixture is separated and the ancillary group iscleaved to provide the pure desired enantiomers. Alternatively, when amolecule contains a basic functional group (such as an amino) or anacidic functional group (such as a carboxyl), it forms a salt ofdiastereomer with a suitable optically active acid or base, and then adiastereomer resolution is performed by a conventional method well knownin the art, followed by recovering to give pure enantiomers. Inaddition, the separation of the enantiomers and diastereomers isgenerally accomplished by the use of chromatography adopting a chiralstationary phase, and optionally in combination with chemicalderivatization method (e.g., forming carbamates from amines).

The compound of the present invention may contain non-naturalproportions of atomic isotopes on one or more atoms which constitute thecompound. For example, the compound may be labeled with a radioisotope,such as tritium (³H), iodine-125 (¹²⁵I) or C-14 (¹⁴C). Any isotopiccomposition transformations of the compound of the present invention,whether are radioactive or not, are included in the scope of the presentinvention.

The term “pharmaceutically acceptable carrier” refers to any formulationor carrier medium capable of delivering an effective amount of theactive substance of the present invention, without interfering with thebiological activity of the active substance and having no toxic sideeffects on the host or patient. Representative carriers include water,oils, vegetables and minerals, cream bases, lotion bases, ointmentbases, etc. These bases include suspensions, tackifiers, transdermalenhancers, etc. Their formulations are well known to the skilled in thecosmetic field or topical drug field. Other information about carrierscan refer to Remington: The Science and Practice of Pharmacy, 21^(st)Ed., Lippincott, Williams & Wilkins (2005), the contents of which areincorporated herein by reference.

The term “excipient” generally refers to the carrier, diluent and/ormedium which is required to formulate an effective pharmaceuticalcomposition.

For drugs or pharmacologically active agents, the term “effectiveamount” or “therapeutically effective amount” refers to a sufficientamount of a drug or agent that is non-toxic but can achieve the desiredeffect.

For the oral dosage form of the present invention, the “effectiveamount” of one active substance in the composition means the amountneeded to achieve the desired effect when used in combination withanother active substance in the composition. The determination of theeffective amount varies with each individual, depending on the age andgeneral condition of the subject, as well as the specific activesubstance. The appropriate effective amount in each case can bedetermined by the skilled in the art according to a routine experiment.

The term “active ingredient”, “therapeutic agent”, “active substance” or“active agent” refers to a chemical entity that can effectively treattarget disorders, diseases or conditions.

When the number of a linking group is 0, such as —(CRR)₀—, it means thatthe linking group is a single bond. When a substituent is vacant, itmeans that the substituent does not exist. For example, when X is vacantin A-X, the structure is actually A. When a bond of one substituent cancross-link to two atoms on one ring, this substituent may be bonded toany atom on the ring. When it does not specify through which atom thelisted substituent is linked to a compound included but not specificallymentioned in a chemical structure formula, this substituent may bebonded through any of its atoms. The combination of substituents and/orvariants thereof is allowable only if such combination will result instable compounds. For example, the structural unit

or indicates that a substitution may occur at any position on cyclohexylor cyclohexadiene.

Unless otherwise defined, the term “halogenated element” or “halogen”per se or as a part of another substituent denotes a fluorine, chlorine,bromine or iodine atom. Furthermore, the term “haloalkyl” is intended toinclude monohaloalkyl and polyhaloalkyl. For example, the term“halo(C₁-C₄)alkyl” is intended to include, but is not limited to,trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl,etc. Examples of haloalkyl include, but are not limited to,trifluoromethyl, trichloromethyl, pentafluoroethyl, andpentachloroethyl. “Alkoxy” represents the above alkyl having a specificnumber of carbon atoms linked through an oxygen bridge. C₁₋₆ alkoxyincludes C₁, C₂, C₃, C₄, C₅ and C₆ alkoxy. Examples of alkoxy include,but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy,n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy and S-pentoxy. “Cycloalkyl”includes saturated ring groups, such as cyclopropyl, cyclobutyl orcyclopentyl. 3- to 7-membered cycloalkyl includes C₃, C₄, C₅, C₆ and C₇cycloalkyl. “Alkenyl” includes a linear or branched hydrocarbon chain,in which one or more carbon-carbon double bonds are present at anystable site on the chain, for example, vinyl and propenyl.

The term “protecting group” includes, but is not limited to, “aminoprotecting group”, “hydroxy protecting group” or “thiol protectinggroup”. The term “amino protecting group” refers to a protecting groupsuitable for preventing side reactions on the nitrogen position of theamino. Representative amino protecting groups include, but are notlimited to: formyl; acyl, such as alkanoyl (e.g., acetyl,trichloroacetyl or trifluoroacetyl); alkoxycarbonyl, such astert-butoxycarbonyl (Boc); arylmethoxycarbonyl, such asbenzyloxycarbonyl (Cbz) and 9-fluorenylmethyloxycarbonyl (Fmoc);arylmethyl, such as benzyl (Bn), trityl (Tr),1,1-di-(4′-methoxyphenyl)methyl; silyl, such as trimethylsilyl (TMS) andtert-butyldimethylsilyl (TBS), etc. The term “hydroxy protecting group”refers to protecting groups suitable for preventing side reactions ofhydroxyl. Representative hydroxy protecting groups include, but are notlimited to: alkyl, such as methyl, ethyl, and tert-butyl; acyl, such asalkanoyl (e.g., acetyl); arylmethyl, such as benzyl (Bn),p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl(benzhydryl, DPM); silyl, such as trimethylsilyl (TMS),tert-butyldimethylsilyl (TBS), etc. An important consideration factor inany synthetic route planning in the art is the selection of suitableprotecting groups for reactive functional groups (such as amino in thepresent application). For trained practitioners, (Protective Groups InOrganic Synthesis, Wiley and Sons, 1991) of Greene and Wuts isauthoritative in this regard. All references cited in the presentapplication are incorporated herein in their entirety.

The compound of the present invention can be prepared by a variety ofsynthetic methods well known by the skilled in the art, including thefollowing exemplified embodiments, the embodiments formed by combiningthem with other chemical synthesis methods, and equivalent alternativesknown to the skilled in the art. Preferred embodiments include, but arenot limited to the examples of the present invention.

The solvents used in the present invention are commercially available.The present invention employs the following abbreviations: aq representsaqua; HATU representsO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate; EDC representsN-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride; m-CPBArepresents 3-chloroperoxybenzoic acid; eq represents equivalent, equalquantity; CDI represents carbonyl diimidazole; DCM representsdichloromethane; PE represents petroleum ether; DIAD representsdiisopropyl azodicarboxylate; DMF represents N,N-dimethylformamide; DMSOrepresents dimethylsulfoxide; EtOAc represents ethyl acetate; EtOHrepresents ethanol; MeOH represents methanol; CBz representscarbobenzyloxy, which is an amine protecting group; BOC representst-butyl carbonyl, which is an amine protecting group; HOAc representsacetic acid; NaCNBH₃ represents sodium cyanoborohydride; r.t. representsroom temperature; O/N represents overnight; THF representstetrahydrofuran; Boc₂O represents di-tert-butyl dicarbonate; TFArepresents trifluoroacetic acid; DIPEA represents diisopropylethylamine;SOCl₂ represents thionyl chloride; CS₂ represents carbon disulfide; TsOHrepresents p-toluenesulfonic acid; NFSI representsN-fluoro-N-(phenylsulfonyl)benzenesulfonamide; NCS represents1-chloropyrrolidine-2,5-dione; n-Bu₄NF represents tetrabutylammoniumfluoride; iPrOH represents 2-propanol; mp represents melting point; LDArepresents lithium diisopropylamide.

DETAILED DESCRIPTION OF THE INVENTION

In order to illustrate the present invention in more details, thefollowing examples are given, but the scope of the present invention isnot limited thereto.

Reference Example 1: Preparation of INT-747

Reference Example 1A

To a solution of chenodeoxycholic acid (60.0 g, 152.8 mmol) inmethanol/acetic acid/water/ethyl acetate (360/120/30/780 mL),tetrabutylammonium bromide (81.0 g, 251.3 mmol) and sodium bromide (9.0g, 87.5 mmol) were added in portions, and then sodium hypochlorite (210mL, 3.4 mol) was added dropwise at 0° C. within 30 min. After beingstirred at 28° C. for 16 hours, saturated sodium bisulfite solution (500mL) was added for quenching. The aqueous layer was extracted with ethylacetate (1000 mL×2), and the combined organic layer was washed withwater (1000 mL×5). The organic layer was dried over sodium sulfate,filtered and evaporated to dryness. The residue was recrystallized(dichloromethane, 200 mL) to give Reference Example 1A (yellow solid, 41g, 69% yield). ¹H NMR (400 MHz, METHANOL-d₄) δ 3.44-3.60 (m, 1H), 2.99(dd, J=5.77, 12.30 Hz, 1H), 2.54 (t, J=11.29 Hz, 1H), 2.08-2.41 (m, 3H),2.00-2.08 (m, 1H), 1.75-1.96 (m, 6H), 1.28-1.69 (m, 9H), 1.09-1.27 (m,8H), 0.97 (d, J=6.53 Hz, 3H), 0.71 (s, 3H).

Reference Example 1B

To a solution of Reference Example 1A (246.0 g, 629.9 mmol) in methanol(2 L), p-toluenesulfonic acid (10.9 g, 63.0 mmol) was added in oneportion, and they were reacted at 80° C. for 4 hours. After being cooledto room temperature, they were evaporated to dryness, and quenched withsaturated sodium bicarbonate solution (1500 mL). Then, the aqueous layerwas extracted with ethyl acetate (1500 mL×3). The combined organic layerwas washed with brine (1000 mL×3), dried over sodium sulfate, filteredand evaporated to dryness. The residue was recrystallized (ethylacetate, 500 mL) to give Reference Example 1 B (white solid, 202 g, 79%yield). ¹H NMR (400 MHz, CHLOROFORM-d) δ 3.66 (s, 3H), 3.55-3.62 (m,1H), 2.85 (dd, J=6.02, 12.55 Hz, 1H), 2.28-2.43 (m, 2H), 2.13-2.26 (m,2H), 1.64-2.04 (m, 10H), 1.19-1.51 (m, 11H), 1.00-1.17 (m, 3H),0.86-0.97 (m, 3H), 0.65 (s, 3H).

Reference Example 1C

To a solution of chlorotrimethylsilane (107.5 g, 989.5 mmol) intetrahydrofuran (500 mL), lithium diisopropylamide (87.4 g, 815.6 mmol)was added dropwise at −78° C. under nitrogen atmosphere, and after beingstirred for 40 min, a solution of Reference Example 1B (50 g, 123.6mmol) in tetrahydrofuran (300 mL) was further added dropwise. Aftercompletion of the dropwise addition, they were stirred at −78° C. foranother 40 min, and then triethylamine (182.5 g, 1.8 mol) was added.They were quenched with saturated sodium hydrogencarbonate (1000 mL)after 1 hour, and the aqueous layer was extracted with ethyl acetate(1000 mL×3). The combined organic layer was further washed with water(100 mL×6) and saturated saline solution (1000 mL×2). The organic layerwas dried over sodium sulfate, filtered and evaporated to dryness, so asto give Reference Example 1C (brown-yellow oil, 68 g, 100% yield), whichcan be directly used in the next step without further purification. ¹HNMR (400 MHz, CHLOROFORM-d) δ 4.75 (dd, J=1.38, 5.90 Hz, 1H), 3.69 (s,3H), 3.48-3.59 (m, 1H), 2.13-2.42 (m, 2H), 1.52-2.04 (m, 1 OH),1.29-1.48 (m, 7H), 0.99-1.23 (m, 5H), 0.95 (d, J=6.53 Hz, 3H), 0.85 (s,3H), 0.70 (s, 3H), 0.17-0.20 (m, 9H), 0.13 (s, 9H).

Reference Example 1D

To a solution of Reference Example 1C (68.0 g, 123.9 mmol) indichloromethane (500 mL), anhydrous acetaldehyde (10.1 g, 229.2 mmol)was added. A solution of boron trifluoride-diethyl ether (64.4 g, 453.4mmol) in dichloromethane (300 mL) was added dropwise at −78° C. undernitrogen atmosphere. The dropping was required to be kept at an internaltemperature of −78° C. After being stirred for 1 hour, it was warmed upto 30° C. and stirred for another 2 hours. The above solution wasquenched with saturated sodium hydrogencarbonate (1000 mL). The aqueouslayer was extracted with dichloromethane (1000 mL×3). The combinedorganic layer was washed with saturated saline solution (1000 mL×2),dried over sodium sulfate, filtered and evaporated to dryness. Theresidue was purified by column chromatography to give Reference Example1D (yellow solid, 43 g, 81% yield). ¹H NMR (400 MHz, CHLOROFORM-d) δ6.12 (q, J=7.03 Hz, 1H), 3.52-3.66 (m, 4H), 2.54 (dd, J=4.02, 13.05 Hz,1H), 2.13-2.40 (m, 5H), 1.68-1.98 (m, 7H), 1.65 (d, J=7.03 Hz, 3H),1.00-1.52 (m, 11H), 0.97 (s, 3H), 0.89 (d, J=6.53 Hz, 3H), 0.61 (s, 3H).

Reference Example 1E

To a solution of Reference Example 1D (212.0 g, 492.3 mmol) in methanol(500 mL), a solution of NaOH (39.4 g, 984.6 mmol) in water (50 mL) wasadded, and they were stirred at 50° C. for 2 hours. After the solventwas evaporated to dryness, water (500 mL) was added, and ethyl acetate(500 mL×2) was employed for extracting. The aqueous phase was adjustedto pH 3 with dilute HCl, and extracted with dichloromethane (600 mL×2).The combined organic layer was concentrated. The residue was purified byrecrystallization (ethanol, 200 mL) to give Reference Example 1E (yellowsolid, 147 g, 72% yield). ¹H NMR (400 MHz, CHLOROFORM-d) δ 6.19 (q,J=7.36 Hz, 1H), 3.60-3.74 (m, 1H), 2.58 (dd, J=4.02, 13.05 Hz, 1H), 2.40(tt, J=5.02, 10.29 Hz, 3H), 2.19-2.32 (m, 2H), 1.61-2.06 (m, 10H),1.04-1.54 (m, 14H), 1.01 (s, 3H), 0.95 (d, J=6.53 Hz, 3H), 0.65 (s, 3H).

Reference Example 1F

To a solution of Reference Example 1E (140.0 g, 336.1 mmol) in asolution of NaOH (0.5 mol) in water (600 mL), 10% Pd—C(19.9 g, 134.4mmol) was added in one portion, and 15 psi of hydrogen was introduced,and they were reacted at 100° C. for 16 hours. After suction filtration,the filtrate was adjusted to pH 3 with dilute hydrochloric acid. Theaqueous layer was extracted with dichloromethane (1500 mL×3). Thecombined organic layer was washed with brine (1000 mL×3), and dried oversodium sulfate, filtered and evaporated to dryness, so as to giveReference Example 1F (white solid, 101 g, 72% yield), which can bedirectly used in the next step without further purification. ¹H NMR (400MHz, CHLOROFORM-d) δ 3.49-3.60 (m, 1H), 2.70 (q, J=6.02 Hz, 1H),2.12-2.45 (m, 4H), 1.65-2.02 (m, 9H), 1.29-1.52 (m, 6H), 1.05-1.24 (m,8H), 0.93 (d, J=6.53 Hz, 5H), 0.81 (t, J=7.53 Hz, 3H), 0.66 (s, 3H).

Reference Compound INT-747

To a solution of Reference Example 1F (16.0 g, 38.2 mmol) in sodiumhydroxide (2 mol, 100 mL), sodium borohydride (8.7 g, 229.3 mmol) wasadded in portions, and they were stirred at 100° C. for 2 hours. Afterbeing cooled to room temperature, saturated aqueous ammonium chloridesolution (150 mL) was added.

The pH thereof was adjusted to 3 with dilute hydrochloric acid. Theaqueous layer was extracted with dichloromethane (300 mL×3). Thecombined organic layer were washed with brine (200 mL×3), dried oversodium sulfate, filtered and evaporated. The residue was purified bycolumn chromatography, so as to give the reference compound INT-747(white solid, 14.5 g, 90% yield). ¹H NMR (400 MHz, CHLOROFORM-d) δ 3.71(br. s, 1H), 3.36-3.48 (m, 1H), 2.18-2.47 (m, 2H), 1.56-2.01 (m, 10H),1.06-1.54 (m, 15H), 0.86-0.97 (m, 9H), 0.66 (s, 3H).

Route 1

Example 1

Example 1A

To a solution of the reference compound INT-747 (2.7 g, 6.4 mmol) andformic acid (0.3 g, 6.4 mmol) in tetrahydrofuran (40 mL), perchloricacid (6.0 g, 60.0 mmol) was added under nitrogen atmosphere. After beingstirred at 55° C. for 6 hours and concentrated under vacuum, the abovesolution was diluted with water (100 mL). The aqueous layer wasextracted with ethyl acetate (100 mL×2). The combined organic layer wasdried over anhydrous sodium sulfate, filtered and evaporated to dryness.The residue was purified by column chromatography, so as to give Example1A as a white solid (2.8 g, 92% yield). ¹H NMR (400 MHz, CHLOROFORM-d) δ8.16 (s, 1H), 8.05 (s, 1H), 5.20 (br. s., 1H), 4.77-4.65 (m, 1H),2.45-2.34 (m, 1H), 2.31-2.21 (m, 1H), 2.01-1.58 (m, 11H), 1.55-1.30 (m,8H), 1.22-1.05 (m, 6H), 0.97-0.88 (m, 9H), 0.66 (s, 3H).

Example 1B

To a solution of Example 1A (100.0 mg, 0.2 mmol) and lead acetate (186.0mg, 0.4 mmol) in carbon tetrachloride (2 mL), iodine (106.0 mg, 0.4mmol) was added, and the reaction system was reacted for 12 hours underlight irradiation. The reaction of the above solution was quenched byadding a solution of sodium thiosulfate (1 mL). The aqueous layer wasextracted with dichloromethane (10 mL×3), dried over anhydrous sodiumsulfate, filtered and evaporated to dryness. The residue was purified bya preparative thin layer plate (petroleum ether:ethyl acetate=5:1), soas to give Example 1B (colorless solid, 50.0 mg, 38% yield). ¹H-NMR(CDCl₃, 400 MHz) δ 8.14 (s, 1H), 8.03 (s, 1H), 5.19 (br. s., 1H),4.62-4.77 (m, 1H), 3.23-3.32 (m, 1H), 3.00-3.12 (m, 1H), 1.96-2.02 (m,2H), 1.85-1.92 (m, 2H), 1.70-1.82 (m, 7H), 1.66-1.72 (m, 2H), 1.39-1.45(m, 2H), 1.23-1.32 (m, 5H), 1.09-1.19 (m, 6H), 0.96 (s, 3H), 0.90-0.93(m, 6H), 0.67 (s, 3H).

Example 1

To a solution of methyl 6-oxopiperidine-3-carboxylate (21.0 mg, 134μmol) in N,N-dimethylformamide (1 mL), sodium hydrogen (7.0 mg, 179μmol, 60%) was added. After half an hour, a solution of Example 1B (50.0mg, 89.5 μmol) in N,N-dimethylformamide (1 mL) was slowly added dropwiseat 0° C. After the dropwise addition was completed, the reaction systemwas slowly warmed up to room temperature, and reacted for 12 hours.Water (3 mL) and lithium hydroxide monohydrate (4 mg, 89.5 mmol) wereadded to the reaction system, and it was stirred for another 3 hours.Water (10 mL) was added, and the reaction system was adjusted to pH=6with hydrochloric acid (1M), extracted withdichloromethane:methanol=10:1 (10 mL×3). The organic layer was driedover anhydrous sodium sulfate, filtered, and evaporated to dryness. Theresidue was purified by preparative thin layer plate(dichloromethane:methanol=10:1), so as to give Example 1 (15 mg, 32.4%yield). ¹H NMR (400 MHz, METHANOL-d₄) δ 3.67 (br. s., 1H), 3.58-3.48 (m,2H), 3.45-3.34 (m, 2H), 3.31-3.27 (m, 1H), 2.87 (d, J=13.1 Hz, 1H),2.49-2.29 (m, 2H), 2.11 (br. s., 2H), 2.00-1.35 (m, 19H), 1.30-1.10 (m,6H), 1.04 (d, J=6.5 Hz, 3H), 0.95-0.90 (m, 6H), 0.72 (s, 3H).

Example compounds 2-16 were synthesized via the same procedure asExample 1, and shown as follows:

Compound No. Yield % Compound structure ¹H NMR Example 2   6%

¹H NMR (400 MHz, CHLORO- FORM-d) δ 3.97-3.88 (m, 1H), 3.77-3.66 (m, 2H),3.47-3.37 (m, 1H), 3.17-3.04 (m, 1H), 2.83 (q, J = 9.1 Hz, 1H),2.37-2.31 (m, 2H), 2.08 (d, J = 12.8 Hz, 7H), 1.93-1.79 (m, 4H),1.69-1.45 (m, 7H), 1.23-1.11 (m, 4H), 1.00 (d, J = 4.5 Hz, 3H),0.92-0.90 (m, 8H), 0.88 (d, J = 2.0 Hz, 4H), 0.68 (s, 3H) Example 3   6%

¹H NMR (400 MHz, CHLORO- FORM-d) δ 4.03-3.91 (m, 1H), 3.73 (t, J = 7.2Hz, 1H), 3.68 (br. s., 1H), 3.41 (d, J = 4.8 Hz, 1H), 3.25-3.16 (m, 1H),3.05 (br. s., 1H), 2.87-2.80 (m, 1H), 2.39- 2.25 (m, 2H), 2.06-1.95 (m,2H), 1.92-1.77 (m, 6H), 1.77-1.76 (m, 1H), 1.72-1.56 (m, 4H), 1.50 (d, J= 5.0 Hz, 3H), 1.44 (d, J = 12.3 Hz, 3H), 1.19-1.11 (m, 4H), 1.06- 1.00(m, 1H), 0.96 (d, J = 4.5 Hz, 3H), 0.91-0.89 (m, 1H), 0.90 (br. s., 3H),0.88 (br. s., 2H), 0.86 (d, J = 2.5 Hz, 4H), 0.84 (d, J = 3.0 Hz, 2H),0.68-0.62 (m, 3H) Example 4 52.7%

¹H NMR (400 MHz, CHLORO- FORM-d) δ 7.96 (s, 1H), 7.93 (s, 1H), 4.26-4.16(m, 1H), 4.15- 4.06 (m, 1H), 3.70 (br. s., 1H), 3.46-3.33 (m, 1H),2.10-2.02 (m, 1H), 1.99-1.86 (m, 2H), 1.84-1.77 (m, 3H), 1.71-1.56 (m,4H), 1.50- 1.40 (m, 5H), 1.34-1.30 (m, 2H), 1.26 (s, 3H), 1.22-1.17 (m,3H), 1.02 (d, J = 6.5 Hz, 4H), 0.92 (s, 1H), 0.89 (s, 3H), 0.88-0.85 (m,2H), 0.64 (s, 3H) Example 5  57%

¹H NMR (400 MHz, METHANOL- d₄) δ 8.40 (s, 1H), 7.97 (dd, J = 2.0, 9.5Hz, 1H), 6.54 (d, J = 9.5 Hz, 1H), 4.20-3.98 (m, 2H), 3.67 (br. s., 1H),3.32-3.28 (m, 1H), 2.05-1.73 (m, 8H), 1.63-1.21 (m, 16H), 1.12 (d, J =6.0 Hz, 3H), 1.07-0.99 (m, 1H), 0.95-0.90 (m, 6H), 0.71 (s, 3H). Example6 10.9%

¹H NMR (400 MHz, METHANOL- d₄) δ 7.66 (d, J = 5.0 Hz, 1H), 7.15-6.97 (m,1H), 6.82 (br. s., 1H), 4.17-3.92 (m, 2H), 3.66 (br. s., 1H), 3.32-3.29(m, 1H), 2.04- 1.25 (m, 25H), 1.26-1.25 (m, 1H), 1.11 (d, J = 6.0 Hz,3H), 0.95-0.89 (m, 6H), 0.70 (s, 3H). Example 7  53%

¹H NMR (400 MHz, METHANOL- d₄) δ 0.75 (s, 3 H) 0.87-0.97 (m, 6 H) 1.07(d, J = 6.53 Hz, 3 H) 1.12-2.11 (m, 25 H) 3.27-3.32 (m, 1 H) 3.69 (br.s., 1 H) 4.01-4.17 (m, 2 H) 7.16 (dd, J = 8.16, 2.13 Hz, 1 H) 7.38 (t, J= 7.91 Hz, 1 H) 7.54 (s, 1 H) 7.61 (d, J = 7.53 Hz, 1 H). Example 8  16%

¹H NMR (400 MHz, METHANOL- d₄) δ 0.74 (s, 3 H) 0.90-0.96 (m, 6 H) 1.06(d, J = 6.53 Hz, 3 H) 1.21-2.13 (m, 25 H) 3.31 (br. s., 1 H) 3.68 (br.s., 1 H) 4.07-4.27 (m, 2 H) 7.03 (t, J = 7.53 Hz, 1 H) 7.15 (d, J = 8.53Hz, 1 H) 7.46- 7.66 (m, 1 H) 7.82 (dd, J = 7.78, 1.25 Hz, 1 H). Example9 10.9%

¹H NMR (400 MHz, CD₃OD) δ 8.71 (br. s., 1H), 8.37 (br. s., 1H), 7.89(br. s., 1H), 4.16 (d, J = 6.8 Hz, 2H), 3.67 (br. s., 1H), 3.37 (s, 1H),2.01-1.20 (m, 25H), 1.06 (d, J = 6.5 Hz, 3H), 0.94-0.90 (m, 6H), 0.73(s, 3H). Example 10  16%

¹H NMR (400 MHz, METHANOL- d₄) δ 8.57 (br. s., 1H), 8.47 (d, J = 8.3 Hz,1H), 8.19 (d, J = 8.0 Hz, 1H), 4.37 (d, J = 5.3 Hz, 2H), 3.68 (br. s.,1H), 3.37 (s, 1H), 2.03-1.28 (m, 25H), 1.10 (d, J = 6.3 Hz, 3H),0.95-0.91 (m, 6H), 0.75 (s, 3H). Example 11  72%

¹H NMR (400 MHz, METHANOL- d₄) δ 8.71 (br. s., 1H), 8.37 (br. s., 1H),7.89 (br. s., 1H), 4.16 (d, J = 6.8 Hz, 2H), 3.67 (br. s., 1H), 3.37 (s,1H), 2.01-1.20 (m, 25H), 1.06 (d, J = 6.5 Hz, 3H), 0.94-0.90 (m, 6H),0.73 (s, 3H). Example 12  61%

¹H NMR (400 MHz, METHANOL- d₄) δ 8.62 (s, 2H), 4.38-4.26 (m, 2H), 3.68(br. s., 1H), 3.31-3.25 (m, 1H), 2.13-1.67 (m, 9H), 1.65- 1.14 (m, 15H),1.12-0.98 (m, 4H), 0.96-0.89 (m, 6H), 0.75 (s, 3H). Example 13  31%

¹H NMR (400 MHz, METHANOL- d₄) δ 8.40 (s, 1H), 8.22 (br. s., 1H), 7.52(br. s., 1H), 4.22 (dd, J = 7.5, 15.8 Hz, 2H), 3.68 (br. s., 1H),3.35-3.34 (m, 1H), 2.01-1.37 (m, 25H), 1.06 (d, J = 6.3 Hz, 3H),0.94-0.89 (m, 6H), 0.74 (s, 3H). Example 14  16%

¹H NMR (400 MHz, METHANOL- d₄) δ 3.72-3.63 (m, 3H), 3.54-3.39 (m, 4H),2.26 (s, 3H), 1.96 (s, 3H), 1.88-1.04 (m, 25H), 0.99-0.87 (m, 9H), 0.70(s, 3H) Example 15  75%

¹H NMR (400 MHz, METHANOL- d₄) δ 7.67-7.48 (m, 1H), 7.06-6.94 (m, 1H),4.34-4.12 (m, 2H), 3.66 (br s, 1H), 3.30-3.26 (m, 1H), 2.06- 1.14 (m,25H), 1.04 (d, J = 6.5 Hz, 3H), 0.94-0.86 (m, 6H), 0.72 (s, 3H) Example16  34%

¹H NMR (400 MHz, METHANOL- d₄) δ 8.40 (s, 1H), 8.22 (br s, 1H), 7.52 (brs, 1H), 4.22 (br dd, J = 7.5, 15.8 Hz, 2H), 3.68 (br s, 1H), 3.35- 3.34(m, 1H), 2.01-1.37 (m, 25H), 1.06 (br d, J = 6.3 Hz, 3H), 0.94- 0.89 (m,6H), 0.74 (s, 3H)

Route 2

Example 17 and Example 18

Example 2A

To a solution of Example 1A (500.0 mg, 1 mmol) in tetrahydrofuran (5mL), triethylamine (153.0 mg, 1.5 mmol) and ethyl chloroformate (167.0mg, 1.5 mmol) were added, and the reaction system was reacted at 25° C.for 2 hours. The system was cooled to 0° C., and a solution of sodiumborohydride (210.0 mg, 5.6 mmol) in methanol (5 mL) was slowly added tothe reaction system, and they were reacted at 0° C. for 15 min, andfurther reacted at 25° C. for 15 min. The reaction was quenched with 0.2M dilute hydrochloric acid (1 mL). The aqueous layer was extracted withethyl acetate (10 mL×3). The organic layer was dried over anhydroussodium sulfate, filtered and evaporated to dryness. The residue waspurified by column chromatography (petroleum ether:ethyl acetate=4:1),so as to give Example 2A (250.0 mg, 70%). ¹H NMR (400 MHz, CHLOROFORM-d)δ 8.15 (s, 1H), 8.04 (s, 1H), 5.19 (br. s., 1H), 4.77-4.66 (m, 1H),3.66-3.57 (m, 2H), 2.06-1.77 (m, 7H), 1.45-1.06 (m, 21H), 0.97-0.88 (m,9H), 0.66 (s, 3H).

Example 2B

To a mixed solution of Example 2A (280.0 mg, 605 μmol),triphenylphosphine (476.0 mg, 1.8 mmol) and imidazole (124.0 mg, 1.8mmol) in toluene (4 mL) and acetonitrile (1 mL), iodine (461.0 mg, 1.8mmol) was added, and the reaction system was reacted at 25° C. for 3hours. A solution of saturated sodium sulfite (10 mL) was added to thereaction system, and the aqueous layer was extracted with ethyl acetate(10 mL×3).

The organic layers were combined and dried over anhydrous sodiumsulfate, filtered, and evaporated to dryness. The residue was purifiedby column chromatography (petroleum ether:ethyl acetate=20:1), so as togive Example 2B (250.0 mg, 70%). ¹H-NMR (CDCl₃, 400 MHz) ⋅ δ 8.16 (s,1H), 8.07-8.02 (m, 1H), 5.20 (br. s., 1H), 4.76-4.66 (m, 1H), 3.24-3.08(m, 2H), 2.01-1.72 (m, 1 OH), 1.46-1.06 (m, 17H), 0.97-0.89 (m, 9H),0.66 (s, 3H).

Example 17 and Example 18

To a solution of methyl 6-hydroxynicotinate (80.0 mg, 523 μmol) inN,N-dimethylformamide (3 mL), sodium hydrogen (21.0 mg, 523 mmol, 60%)was added at 0° C. After half an hour, a solution of Example 2B (150.0mg, 262 μmol) in N,N-dimethylformamide (5 mL) was added dropwise at 0°C. After the dropwise addition was completed, the reaction system wasslowly warmed up to room temperature, and reacted for 12 hours. Water (3mL) and lithium hydroxide monohydrate (55 mg, 1.31 mmol) were added tothe reaction system, and stirred for another 3 hours. Water (10 mL) wasadded, and the system was adjusted to pH=6 with hydrochloric acid (1M),extracted with dichloromethane:methanol=10:1 (10 mL×3). The organiclayer was dried over anhydrous sodium sulfate, filtered and evaporatedto dryness. The residue was isolated and purified by a preparative thinlayer plate (dichloromethane:methanol=10:1), so as to give Example 17(40 mg, 29%), ¹H NMR (400 MHz, METHANOL-d₄) δ 8.44 (d, J=2.5 Hz, 1H),7.97 (dd, J=2.5, 9.5 Hz, 1H), 6.55 (d, J=9.5 Hz, 1H), 4.10-3.96 (m, 2H),3.70-3.63 (m, 1H), 3.37-3.34 (m, 1H), 2.02-1.11 (m, 27H), 0.98 (d, J=6.0Hz, 3H), 0.95-0.88 (m, 6H), 0.71 (s, 3H); and Example 18 (20 mg, 14%),¹H NMR (400 MHz, METHANOL-d₄) δ 8.77 (s, 1H), 8.21 (dd, J=1.8, 8.8 Hz,1H), 6.84 (d, J=8.5 Hz, 1H), 4.34 (d, J=2.5 Hz, 2H), 3.78-3.58 (m, 1H),3.38-3.33 (m, 1H), 2.02-1.22 (m, 27H), 1.01 (d, J=6.5 Hz, 3H), 0.95-0.90(m, 6H), 0.72 (s, 3H).

Route 3

Example 19

Example 3A

To a solution of Example 2A (500.0 mg, 1.1 mmol), copper acetate (99.0mg, 0.6 mmol) and lead acetate (2.4 g, 11.0 mmol) in toluene (5 mL),pyridine (980.0 mg, 12.4 mmol) was added, and the reaction system wasreacted at 110° C. for 12 hours. The reaction system was filtered, andthe filtrate was concentrated. The residue was purified by columnchromatography (petroleum ether:ethyl acetate=20:1), so as to give thetitle compound (120.0 mg, 27.0%). ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.15(s, 1H), 8.05 (s, 1H), 5.71-5.59 (m, 1H), 5.19 (br. s., 1H), 4.93-4.80(m, 2H), 4.76-4.67 (m, 1H), 2.12-1.61 (m, 11H), 1.52-1.09 (m, 13H), 1.04(d, J=6.8 Hz, 3H), 0.97 (s, 3H), 0.91 (t, J=7.4 Hz, 3H), 0.68 (s, 3H).

Example 3B

To a solution of Example 3A (100.0 mg, 0.2 mmol), sodium carbonate (74.0mg, 0.7 mmol) and tetrabutylammonium bromide (66.0 mg, 0.2 mmol) inN,N-dimethylformamide (2 mL), palladium acetate (5.0 mg, 23.0 μmol) andmethyl 2-iodobenzoate (60.8 mg, 0.2 mmol) were added under nitrogen, andthe reaction system was reacted at 85° C. for 12 hours. The solvent wasevaporated to dryness and water (10 mL) was added to the reactionsystem. The aqueous layer was extracted with dichloromethane (20 mL×3).The organic layers were combined. The organic phase was dried overanhydrous sodium sulfate, filtered and evaporated to dryness. Theresidue was purified by preparative thin layer chromatography (petroleumether:ethyl acetate=10:1), so as to give the title compound (80.0 mg,61.0%). ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.16 (s, 1H), 8.07 (s, 1H),8.01 (s, 1H), 7.90-7.84 (m, 1H), 7.51 (d, J=7.8 Hz, 1H), 7.41-7.33 (m,1H), 6.40-6.31 (m, 1H), 6.16 (dd, J=8.8, 15.8 Hz, 1H), 5.21 (br. s.,1H), 4.79-4.63 (m, 1H), 3.94 (s, 3H), 2.29 (d, J=6.8 Hz, 1H), 2.08-1.62(m, 11H), 1.52-1.20 (m, 11H), 1.15 (d, J=6.5 Hz, 3H), 1.00 (s, 3H),0.95-0.90 (m, 3H), 0.74 (s, 3H).

Example 3C

To a solution of Example 3B (100.0 mg, 0.2 mmol) in methanol (3 mL), wetpalladium on carbon (50 mg, 10%) was added under nitrogen, and thereaction system was reacted under hydrogen condition (15 psi) at 25° C.for 12 hours. The reaction system was filtered and evaporated todryness, so as to give Example 3C (90.0 mg, 90.0%). ¹H NMR (400 MHz,CHLOROFORM-d) δ 8.18-8.11 (m, 1H), 8.06-8.00 (m, 1H), 7.85-7.81 (m, 2H),7.35-7.31 (m, 2H), 5.18 (br. s., 1H), 4.76-4.51 (m, 1H), 2.77-2.64 (m,1H), 2.54-2.44 (m, 1H), 1.95-1.20 (m, 25H), 1.01 (d, J=6.5 Hz, 3H),0.95-0.87 (m, 6H), 0.64 (s, 3H).

Example 19

To a solution of Example 3C (160.0 mg, 282.0 μmol) in tetrahydrofuran (1mL), methanol (1 mL) and water (1 mL), lithium hydroxide (59.0 mg, 1.4mmol) was added. The reaction system was reacted at 20° C. for 12 hours.Water (5 mL) was added to the reaction system, and the system wasadjusted to pH=1-2 with hydrochloric acid (1 M). The aqueous phase wasextracted with ethyl acetate (10 mL×3). The organic layers were combinedand dried over anhydrous sodium sulfate, filtered and evaporated todryness. The residue was isolated and purified by preparative isolation(HCl), so as to give Example 19 (80 mg, 57%). ¹H NMR (400 MHz,METHANOL-d₄) δ 7.93-7.76 (m, 2H), 7.45-7.35 (m, 2H), 3.67 (br. s., 1H),3.37-3.35 (m, 1H), 2.84-2.73 (m, 1H), 2.60 (d, J=10.5 Hz, 1H), 2.06-1.27(m, 25H), 1.09 (d, J=6.3 Hz, 3H), 0.94-0.91 (m, 6H), 0.70 (s, 3H).

Route 4

Example 20

To a solution of Example 3B (40.0 mg, 70.8 μmol) in tetrahydrofuran (0.5mL), methanol (0.5 mL) and water (0.5 mL), lithium hydroxide (15 mg,0.35 mmol) was added. The reaction system was reacted at 20° C. for 12hours. Water (5 mL) was added to the reaction system, and the system wasadjusted to pH=1-2 with hydrochloric acid (1 M). The aqueous phase wasextracted with ethyl acetate (10 mL×3). The organic layers were combinedand dried over anhydrous sodium sulfate, filtered and evaporated todryness. The residue was purified by preparative thin layerchromatography (dichloromethane:methanol=10:1), so as to give Example 20(25.0 mg, 71.0%). ¹H NMR (400 MHz, METHANOL-d₄) δ 8.00 (br. s., 1H),7.84 (d, J=7.0 Hz, 1H), 7.55 (d, J=7.5 Hz, 1H), 7.42-7.34 (m, 1H),6.46-6.32 (m, 1H), 6.19 (dd, J=8.8, 15.6 Hz, 1H), 3.72-3.62 (m, 1H),3.39-3.37 (m, 1H), 2.32 (d, J=6.0 Hz, 1H), 2.07-1.29 (m, 22H), 1.17 (d,J=6.3 Hz, 3H), 0.95-0.89 (m, 6H), 0.78 (s, 3H).

Route 5

Example 21

Example 5A

After a mixture of Reference Example 1 (100.0 mg, 0.2 mmol),triethylamine (48.0 mg, 0.5 mmol) and O,N-dimethylhydroxylaminehydrochloride (23.0 mg, 0.2 mmol) in acetonitrile (2 mL) were stirred at25° C. for 0.5 hour, O-benzotriazol-N,N,N′,N′-tetramethyluroniumtetrafluoroborate (95.0 mg, 0.3 mmol) was added thereto. The resultingmixture was stirred at 25° C. for 12 hours. After the solvent wasremoved by evaporation under vacuum, the residue was purified by columnchromatography, so as to give Example 5A as a white solid (90 mg, 82%yield). ¹H NMR (400 MHz, CHLOROFORM-d) δ 3.73-3.67 (m, 4H), 3.45-3.36(m, 1H), 3.18 (s, 3H), 2.51-2.40 (m, 1H), 2.38-2.27 (m, 1H), 2.00-1.89(m, 2H), 1.87-1.74 (m, 5H), 1.71-1.56 (m, 5H), 1.53-1.31 (m, 11H),1.23-1.14 (m, 3H), 1.06-0.99 (m, 1H), 0.96 (d, J=6.3 Hz, 3H), 0.93-0.88(m, 6H), 0.67 (s, 3H).

Example 5B

To a solution of Example 5A (100.0 mg, 0.2 mmol) in tetrahydrofuran (5mL), a solution of methylmagnesium bromide (0.4 mL, 1.1 mmol, 3N) indiethyl ether was added at 0° C., and they were stirred at 0° C. foranother 30 min, and then warmed up to room temperature and stirred for12 hours. The reaction was quenched by adding ice water, and then theywere extracted with ethyl acetate (60 mL×2). After being washed withwater, the organic phase was dried over anhydrous sodium sulfate, andfiltered. After the solvent was removed under vacuum, the residue waspurified by preparative TLC, so as to give Example 5B as a white solid(6 mg, 66% yield). ¹H NMR (400 MHz, CHLOROFORM-d) δ 3.71 (br. s., 1H),3.48-3.35 (m, 1H), 2.51-2.41 (m, 1H), 2.39-2.29 (m, 1H), 2.14 (s, 3H),1.99-1.79 (m, 5H), 1.76-1.56 (m, 5H), 1.51-1.30 (m, 10H), 1.22-1.11 (m,3H), 1.00 (dt, J=3.3, 14.2 Hz, 1H), 0.94-0.87 (m, 9H), 0.66 (s, 3H).

Example 5C

Example 5B (1.0 g, 2.4 mmol) was dissolved in 1,4-dioxane (20.0 mL), and1,4-dihydropyran (2.0 g, 23.9 mmol, 2.2 mL) and p-toluenesulfonic acid(90.9 mg, 478.0 μmol) were added thereto. The reaction solution wasstirred at 30° C. for 36 hours. The solvent was removed byconcentration, and water (5 mL) was added to the reaction solution,which was extracted with ethyl acetate (10 mL×3). The organic layer waswashed with water (10 mL), dried over anhydrous sodium sulfate, filteredand concentrated, so as to give a crude product. The crude product wasisolated by column chromatography, so as to give Example 5C (450.0 mg,32.1% yield, colorless oil). ¹H NMR (400 MHz, CHLOROFORM-d) δ 4.73 (d,J=3.3 Hz, 1H), 4.56 (d, J=4.3 Hz, 1H), 3.95-3.81 (m, 2H), 3.76-3.72 (m,1H), 3.55-3.31 (m, 4H), 2.52-2.41 (m, 1H), 2.34 (ddd, J=5.8, 9.9, 16.0Hz, 1H), 1.97-1.80 (m, 7H), 1.74-1.67 (m, 4H), 1.55 (br. s., 4H), 1.41(d, J=7.3 Hz, 3H), 1.33-1.27 (m, 3H), 1.17-1.08 (m, 3H), 0.90 (s, 3H),0.88 (s, 3H), 0.69-0.59 (m, 3H).

Example 5D

Metall sodium (68.2 mg, 3.0 mmol) was dissolved in anhydrous ethanol (5mL), cooled to 0° C., to which Example 5C (170.0 mg, 296.8 μmol,dissolved in 2 mL of anhydrous ethanol) was slowly added, and they werestirred for another half an hour at 0° C. Then, diethyl oxalate (65.0mg, 445.1 μmol, 60.8 μL, dissolved in 1 mL of anhydrous ethanol) wasadded dropwise. After being stirred at 0° C. for half an hour, thereaction solution was warmed up to 25° C. and stirred for another 35hours. Ethanol (6 mL) was added to the reaction solution, and thesolvent was removed by concentration. Ethyl acetate (40 mL) was added,and 2 mol of citric acid aqueous solution was added under stirring in anice bath (to pH=5-6). The aqueous phase was extracted with ethyl acetate(10 mL×2). The organic layer was washed with water (10 mL), salinesolution (10 mL), dried over anhydrous sodium sulfate, filtered andconcentrated, so as to give a crude product. The crude product wasisolated by column chromatography, so as to give Example 5D (90.0 mg,44.2% yield). ¹H NMR (400 MHz, CHLOROFORM-d) δ 4.73 (d, J=3.3 Hz, 1H),4.57 (d, J=4.0 Hz, 1H), 4.36 (q, J=7.0 Hz, 2H), 3.97-3.80 (m, 2H), 3.70(br. s., 1H), 3.52-3.34 (m, 3H), 2.54 (ddd, J=5.1, 10.3, 15.4 Hz, 1H),2.44-2.29 (m, 1H), 1.98-1.90 (m, 2H), 1.82 (br. s., 3H), 1.74-1.67 (m,4H), 1.53 (br. s., 7H), 1.38 (s, 3H), 1.26 (t, J=7.0 Hz, 2H), 1.16 (d,J=9.8 Hz, 2H), 0.95 (d, J=6.5 Hz, 2H), 0.89 (s, 3H), 0.71-0.59 (m, 3H).

Example 5E

Example 5D (90.0 mg, 131.0 μmol) was dissolved in ethanol (3.0 mL), andhydroxylamine hydrochloride (27.3 mg, 393.0 μmol) was added thereto, andthe reaction solution was stirred at 80° C. for 5 hours. The solvent wasremoved by concentration, and water (5 mL) was added to the reactionsolution. The aqueous phase was extracted with ethyl acetate (10 mL×3).The organic layer was washed with water (10 mL), dried over anhydroussodium sulfate, filtered, and the filtrate was evaporated to drynessunder vacuum. Example 5E was obtained without purifying the crudeproduct (86.0 mg, crude product, yellow oil). ¹H NMR (400 MHz,CHLOROFORM-d) δ 6.39 (s, 1H), 4.43 (q, J=7.3 Hz, 2H), 3.72 (br. s., 1H),3.43-3.40 (m, 1H), 2.90-2.64 (m, 2H), 2.42 (t, J=6.9 Hz, 2H), 2.00-1.95(m, 2H), 1.75 (d, J=6.8 Hz, 3H), 1.69 (br. s., 4H), 1.60 (d, J=3.3 Hz,3H), 1.48 (d, J=7.5 Hz, 4H), 1.41 (s, 3H), 1.28 (br. s., 4H), 1.20 (d,J=8.5 Hz, 3H), 0.90 (s, 3H), 0.89-0.88 (m, 1H), 0.65 (s, 3H).

Example 21

Example 5E (115.0 mg, 222.9 μmol) was dissolved in tetrahydrofuran (1.0mL), water (1.0 mL) and methanol (1.0 mL), and lithium hydroxidemonohydrate (93.6 mg, 2.2 mmol) were added thereto. The reactionsolution was stirred at 25° C. for 12 hours. The reaction solution wasacidified with 1 mol of hydrochloric acid (to pH=5-6), and extractedwith ethyl acetate (10 mL×3). The organic layer was washed with water(10 mL), dried over anhydrous sodium sulfate and filtered. The filtratewas evaporated to dryness under vacuum. The crude product was isolatedby thin layer chromatography, so as to give Example 21 (28.0 mg, 25.8%yield). ¹H NMR (400 MHz, METHANOL-d₄) δ 6.41 (s, 1H), 3.67 (br. s., 1H),3.31 (br. s., 1H), 2.94-2.83 (m, 1H), 2.80-2.70 (m, 1H), 2.03 (d, J=12.0Hz, 1H), 1.96-1.83 (m, 5H), 1.80-1.73 (m, 2H), 1.64 (br. s., 1H),1.58-1.50 (m, 5H), 1.48-1.42 (m, 2H), 1.42-1.34 (m, 3H), 1.32-1.27 (m,3H), 1.24 (d, J=9.5 Hz, 2H), 1.05 (d, J=6.0 Hz, 3H), 0.93 (s, 3H),0.92-0.89 (m, 3H), 0.71 (s, 3H).

Route 6

R_(b)=H or X, for example:

Example 22

Example 6A

To a solution of methyl 6-hydroxynicotinate (438.7 mg, 2.9 mmol),Example 1B (800 mg, 1.4 mmol) in chloroform (20 mL), silver carbonate(790 mg, 2.9 mmol) was added. The reaction system was reacted at 60° C.for 72 hours, filtered, and concentrated. The residue was isolated bycolumn (petroleum ether:ethyl acetate=10:1), so as to give the titlecompound 6A (400 mg, 47.9%). ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.82 (d,J=2.3 Hz, 1H), 8.16 (s, 1H), 8.16-8.12 (m, 1H), 8.05 (s, 1H), 6.74 (d,J=8.5 Hz, 1H), 5.21 (br. s., 1H), 4.80-4.61 (m, 1H), 4.48-4.27 (m, 2H),3.91 (s, 3H), 2.02-1.63 (m, 11H), 1.46-1.09 (m, 14H), 1.02 (d, J=6.5 Hz,3H), 0.97 (s, 3H), 0.94-0.89 (m, 3H), 0.68 (s, 3H).

Example 22

To a solution of Example 6A (800 mg, 1.4 mmol) in methanol (10 mL), asolution of 10% sodium hydroxide (volume ratio of methanol to water was1:1) (600 mg, 15 mmol, 6 mL) was added, and the reaction system wasreacted at 70° C. for 2 hours. The solvent was evaporated to dryness.The system was adjusted to pH=1-2 with dilute hydrochloric acid (1M) andextracted with dichloromethane:methanol=10:1 (20 mL×3). The organiclayer was dried over anhydrous sodium sulfate, filtered andconcentrated. The residue was purified by preparative chromatography, soas to give the title compound 22 (460 mg, 64% yield). ¹H NMR (400 MHz,METHANOL-d₄) δ 8.78 (d, J=1.8 Hz, 1H), 8.21 (dd, J=2.3, 8.5 Hz, 1H),6.84 (d, J=8.8 Hz, 1H), 4.56-4.23 (m, 2H), 3.68 (br. s., 1H), 3.35-3.34(m, 1H), 2.04-1.17 (m, 25H), 1.07 (d, J=6.5 Hz, 3H), 0.96-0.89 (m, 6H),0.74 (s, 3H).

The preparation of Examples 23-26 was referred to the procedure of thetitle compound 22.

Compound No. Yield % Compound structure ¹H NMR Example 23 47.2%

¹H NMR (400 MHz, METHANOL-d₄) δ 8.68 (br. s., 1H), 8.24 (s, 1H), 4.61-4.37 (m, 2H), 3.67 (br. s., 1H), 3.37 (s, 1H), 2.04-1.18 (m, 25H), 1.08(d, J = 6.3 Hz, 3H), 0.95-0.89 (m, 6H), 0.73 (s, 3H) Example 24 56.6%

¹H NMR (400 MHz, METHANOL-d₄) δ 8.58 (d, J = 1.8 Hz, 1H), 7.94 (dd, J =1.8, 10.5 Hz, 1H), 4.66-4.37 (m, 2H), 3.74-3.58 (m, 1H), 3.32-3.28 (m,1H), 2.08-1.20 (m, 25H), 1.08 (d, J = 6.5 Hz, 3H), 0.95-0.87 (m, 6H),0.73 (s, 3H) Example 25 56.7%

¹H NMR (400 MHz, METHANOL-d₄) δ = 7.85-7.78 (m, 1H), 7.73 (d, J = 7.3Hz, 1H), 7.00 (d, J = 8.3 Hz, 1H), 4.53- 4.34 (m, 2H), 3.68 (br s, 1H),2.02- 1.10 (m, 25H), 1.08 (br d, J = 6.5 Hz, 3H), 0.97-0.88 (m, 6H),0.74 (s, 3H) Example 26 75.6%

¹H NMR (400 MHz, METHANOL-d₄) δ = 8.30 (d, J = 5.5 Hz, 1H), 7.54 (dd, J= 1.3, 5.5 Hz, 1H), 7.43 (s, 1H), 4.57-4.29 (m, 2H), 3.66 (br s, 1H),2.08-1.10 (m, 25H), 1.06 (d, J = 6.5 Hz, 3H), 0.95-0.87 (m, 6H), 0.73(s, 3H)

Route 7

Example 27

To a solution of Example 22 (100.0 mg, 194.7 μmol), DCC (60.3 mg, 292μmol) and DMAP (23.8 mg, 194.7 μmol) in dichloromethane (5 mL),methanesulfonamide (37 mg, 389.3 μmol) was added. The reaction systemwas reacted at 25° C. for 12 hours. Water (5 mL) was added to thesystem. The system was adjusted to pH=2 with dilute hydrochloric acid (1M), and extracted with dichloromethane (10 mL×3). The organic layer wasdried over anhydrous sodium sulfate, filtered and concentrated. Theresidue was purified by preparative chromatography, so as to giveExample 27 (25 mg, 21.7% yield). ¹H NMR (400 MHz, METHANOL-d₄) δ 8.74(d, J=2.3 Hz, 1H), 8.23 (dd, J=2.3, 8.8 Hz, 1H), 7.06-6.87 (m, 1H),4.57-4.40 (m, 2H), 3.67 (br s, 1H), 3.39 (s, 3H), 2.03-1.27 (m, 25H),1.07 (br d, J=6.5 Hz, 3H), 0.96-0.90 (m, 6H), 0.73 (s, 3H).

The preparation of Examples 28-34 was referred to the procedure of thetitle compound 27.

Compound No. Yield % Compound structure ¹H NMR Example 28 50.0

¹H NMR (400 MHz, METHANOL-d₄) δ 8.72 (d, J = 2.3 Hz, 1H), 8.23 (dd, J =2.4, 8.9 Hz, 1H), 6.97 (d, J = 8.8 Hz, 1H), 4.54-4.32 (m, 2H), 3.65 (brs, 1H), 3.56-3.38 (m, 1H), 3.18-3.07 (m, 1H), 2.02-1.15 (m, 29H), 1.05(d, J = 6.3 Hz, 3H), 0.93- 0.87 (m, 6H), 0.71 (s, 3H) Example 29 26.0

¹H NMR (400 MHz, METHANOL-d₄) δ 8.69 (d, J = 2.3 Hz, 1H), 8.16 (dd, J =2.4, 8.9 Hz, 1H), 6.91 (d, J = 8.8 Hz, 1H), 4.56-4.27 (m, 2H), 3.66 (brs, 1H), 3.46 (s, 1H), 2.05-1.57 (m, 15H), 1.50 (s, 9H), 1.33-1.09 (m,10H), 1.05 (d, J = 6.5 Hz, 3H), 0.93-0.86 (m, 6H), 0.71 (s, 3H) Example30 39.8

¹H NMR (400 MHz, METHANOL-d₄) δ = 8.70 (d, J = 2.0 Hz, 1H), 8.13 (dd, J= 2.5, 8.8 Hz, 1H), 6.86 (d, J = 8.8 Hz, 1H), 4.50-4.33 (m, 2H), 3.91(quin, J = 6.9 Hz, 1H), 3.66 (br s, 1H), 3.50- 3.34 (m, 1H), 2.05-1.49(m, 16H), 1.42 (d, J =7.0 Hz, 6H), 1.39-1.08 (m, 9H), 1.05 (d, J = 6.5Hz, 3H), 0.93-0.86 (m, 6H), 0.72 (s, 3H) Example 31 68.5

¹H NMR (400 MHz, CHLOROFORM-d) δ 8.62 (br s, 1H), 8.09 (br d, J = 7.8Hz, 2H), 7.96 (br d, J = 6.0 Hz, 1H), 7.62-7.55 (m, 1H), 7.53- 7.46 (m,2H), 6.78-6.63 (m, 1H), 6.70 (br d, J = 5.8 Hz, 1H), 4.41-4.21 (m, 2H),3.64 (br s, 1H), 3.37 (br s, 1H), 1.98-1.66 (m, 12H), 1.63-1.48 (m, 5H),1.45-1.02 (m, 15H), 0.97-0.89 (m, 4H), 0.85- 0.78 (m, 6H), 0.59 (s, 3H).Example 32 76.8

¹H NMR (400 MHz, CHLOROFORM-d) δ 8.74 (br s, 1H), 8.19- 8.01 (m, 1H),8.12 (br s, 1H), 7.60 (br d, J = 7.8 Hz, 2H), 7.35-7.26 (m, 1H), 7.30(br t, J = 7.5 Hz, 1H), 7.13-7.04 (m, 1H), 6.80 (br s, 1H), 4.46-4.23(m, 2H), 3.64 (br s, 1H), 3.34 (br s, 1H), 1.97-1.68 (m, 15H), 1.63-1.49(m, 5H), 1.46- 1.03 (m, 15H), 1.00-0.88 (m, 4H), 0.86-0.79 (m, 7H), 0.62(s, 3H). Example 33 69.6

¹H NMR (400 MHz, METHANOL-d₄) δ 8.62 (s, 1H), 8.21-8.01 (m, 1H), 6.89(br s, 1H), 4.52-4.30 (m, 2H), 3.68 (br s, 1H), 3.57-3.44 (m, 1H), 2.93(s, 3H), 2.04- 1.25 (m, 25H), 1.07 (d, J = 6.5 Hz, 3H), 0.95- 0.94 (m,1H), 0.95-0.90 (m, 6H), 0.74 (s, 3H) Example 34

¹H NMR (400 MHz, CHLOROFORM-d) δ 9.12 (br s, 1H), 8.63- 8.46 (m, 1H),8.56 (br s, 1H), 8.03 (br s, 1H), 7.17-6.99 (m, 1H), 4.69- 4.38 (m, 2H),3.65-3.28 (m, 5H), 3.03-2.87 (m, 1H), 2.09-1.57 (m, 13H), 1.52-1.13 (m,13H), 1.08-0.96 (m, 4H), 0.93- 0.76 (m, 11H), 0.69 (s, 3H).

Route 8

Example 35

Example 35 was synthesized by using Example 11 as a raw material, andthe procedure was referred to the synthesis of Example 27. The yield was17.4%. ¹H NMR (400 MHz, METHANOL-d₄) δ 8.95-8.70 (m, 2H), 8.48 (s, 1H),4.41-4.27 (m, 2H), 3.67 (br s, 1H), 3.41 (s, 3H), 3.33-3.32 (m, 1H),2.02-1.16 (m, 25H), 1.08 (d, J=6.3 Hz, 3H), 0.94-0.88 (m, 6H), 0.74 (s,3H).

Route 9

Example 36

Example 36 was synthesized by using Example 23 as a raw material, andthe procedure was referred to the synthesis of Example 27. The yield was17.5%. ¹H NMR (400 MHz, CHLOROFORM-d) δ=8.60 (d, J=2.3 Hz, 1H), 8.18 (d,J=2.3 Hz, 1H), 4.56-4.33 (m, 2H), 3.69 (br s, 1H), 3.45 (s, 3H),1.96-1.15 (m, 25H), 1.01 (d, J=6.5 Hz, 3H), 0.89-0.81 (m, 6H), 0.66 (s,3H).

Route 10

Example 37

Example 37 was synthesized by using Example 23 as a raw material, andthe procedure was referred to the synthesis of Example 27. The yield was60.8%. ¹H NMR (400 MHz, METHANOL-d₄) δ 8.55 (d, J=1.8 Hz, 1H), 8.17 (d,J=1.8 Hz, 1H), 4.57-4.38 (m, 2H), 3.87-3.75 (m, 2H), 3.66 (br s, 1H),3.32-3.20 (m, 1H), 3.11 (t, J=6.9 Hz, 2H), 2.06-1.23 (m, 25H), 1.06 (brd, J=6.5 Hz, 3H), 0.96-0.86 (m, 6H), 0.71 (s, 3H).

Route 11

Example 38

Example 38 was synthesized by using Example 22 as a raw material, andthe procedure was referred to the synthesis of Example 27. The yield was53%. ¹H NMR (400 MHz, METHANOL-d₄) δ 8.63-8.62 (m, 1H), 8.09-8.06 (m,1H), 6.84-6.82 (m, 1H), 4.39-4.28 (m, 2H), 3.82-3.74 (m, 3H), 3.40-3.30(m, 1H), 3.12-3.08 (m, 2H), 1.99-1.20 (m, 25H), 1.06 (d, J=6.5 Hz, 3H),0.94-0.90 (m, 6H), 0.73 (s, 3H).

Route 12

Example 39

Example 12A

Glycine hydrochloride (14 mg, 112 μmol) was reacted with a solution oftriethylamine (20.0 μL, 146 μmol) in ethyl acetate (5 mL) at 20° C. for0.5 hour. Example 22 (50 mg, 97 μmol) and2-ethoxy-1-ethoxycarbonyl-1.2-dihydroquinoline (36 mg, 146 μmol) wereadded to the system. The reaction system was reacted at 65° C. for 10hours. Water (5 mL) was added to the system, and the aqueous phase wasextracted with ethyl acetate (15 mL×2). The organic phases were combinedand sequentially washed with 0.5 N of aqueous sodium hydroxide solution(15 mL), water (15 mL), 0.5 N of hydrochloric acid (15 mL) and water (15mL). The organic layer was dried over anhydrous sodium sulfate, filteredand evaporated to dryness. The crude product was isolated and purifiedby preparative isolation (TFA), so as to give Example 12A (25 mg,43.9%). ¹H NMR (400 MHz, CDCl₃) 8.66-8.65 (m, 1H), 8.08-8.05 (m, 1H),6.81-6.67 (m, 1H), 6.68 (s, 1H), 4.42-4.24 (m, 2H), 4.35-4.24 (m, 2H),3.81 (s, 3H), 3.72-3.71 (m, 1H), 3.46-3.44 (m, 1H), 1.99-1.20 (m, 25H),1.03 (d, J=6.5 Hz, 3H), 0.92-0.88 (m, 6H), 0.68 (s, 3H).

Example 39

To a solution of Example 12A (25.0 mg, 42.7 μmol) in tetrahydrofuran (3mL), methanol (1 mL) and water (2 mL), lithium hydroxide (8.9 mg, 213.7μmol) was added. The reaction system was reacted at 30° C. for 4 hours.The system was adjusted to pH=5 with hydrochloric acid (1M), and theaqueous phase was extracted with ethyl acetate (20 mL×2). The organiclayers were combined, and dried over sodium sulfate, filtered andevaporated to dryness. The residue was isolated by preparative TLC, soas to give Example 39 (24 mg, 98%). ¹H NMR (400 MHz, METHANOL-d₄)8.66-8.65 (m, 1H), 8.11-8.02 (m, 1H), 6.84-6.82 (m, 1H), 4.48-4.29 (m,2H), 4.09-4.08 (m, 3H), 3.66-3.65 (m, 1H), 1.99-1.20 (m, 25H), 1.06 (d,J=6.5 Hz, 3H), 0.92-0.88 (m, 6H), 0.72 (s, 3H).

Route 13

Example 40

Example 13A

Using Example 2B as a raw material, the procedure of Example 13A wasreferred to the synthesis of Example 6A. The yield was 16.3%. ¹H NMR(400 MHz, CHLOROFORM-d) δ 8.69 (d, J=2.0 Hz, 1H), 8.21 (d, J=2.0 Hz,1H), 8.16 (s, 1H), 8.05 (s, 1H), 5.20 (br s, 1H), 4.72 (br s, 1H),4.46-4.36 (m, 2H), 3.92 (s, 3H), 2.06-1.66 (m, 15H), 1.31-1.04 (m, 10H),0.97-0.81 (m, 9H), 0.67 (s, 3H).

Example 40

The procedure of Example 40 was referred to the synthesis of Example 22.The yield was 96.4%. ¹H NMR (400 MHz, METHANOL-d₄) δ=8.74 (d, J=2.0 Hz,1H), 8.29 (d, J=1.8 Hz, 1H), 4.51 (t, J=6.4 Hz, 2H), 3.72 (br s, 1H),3.43-3.39 (m, 1H), 2.11-1.23 (m, 27H), 1.07 (d, J=6.5 Hz, 3H), 1.00-0.93(m, 6H), 0.77 (s, 3H).

Route 14

Example 14A

To a solution of Reference Example 1 (40.0 g, 95.1 mmol) in methanol(500 mL), p-toluenesulfonic acid (4.0 g, 21.0 mmol) was added. Thereaction system was reacted at 70° C. for 3 hours. The solvent wasevaporated to dryness, and the system was diluted with dichloromethane(500 mL). The organic phase was sequentially washed with saturatedaqueous solution of sodium carbonate (3×100 mL), aqueous solution (100mL) and saturated saline solution (100 mL), and dried over anhydroussodium sulfate, filtered and concentrated. Example 14A (41 g, 100%yield) was obtained without purification, and can be directly used inthe next step.

Example 14B

Example 14A (41 g, 95.1 mmol) was dissolved in anhydrous tetrahydrofuransolution (300 mL), and warmed up to 50° C. under nitrogen atmosphere,and a solution of methylmagnesium bromide (800 mL, 1 M) intetrahydrofuran was slowly added dropwise. After the dropwise additionwas completed, the reaction solution was cooled to room temperature, andstirred overnight. After the reaction was completed, cyclohexane (25 mL)was added and filtered. The filter residue was dissolved in a mixedsolution of 3N aqueous hydrochloric acid solution (800 mL) anddichloromethane (200 mL). They were stirred for another 30 min. Afterthe reaction was completed, the organic phase was separated from theaqueous phase. The aqueous phase was further extracted withdichloromethane (2×200 mL). The organic phases were combined andsequentially washed with water (100 mL) and saturated saline solution(100 mL), dried over anhydrous sodium sulfate, filtered andconcentrated. Example 14B (51 g, 100% yield) was obtained withoutpurification, and can be directly used in the next step.

Example 14C

Acetic anhydride (9.9 mL, 105.1 mmol), pyridine (1.6 mL, 19.8 mmol) and4-dimethylaminopyridine (0.8 g, 6.6 mmol) were added to a solution ofExample 14B (51 g, 95.1 mmol) in anhydrous tetrahydrofuran (300 mL).

The reaction solution was stirred overnight at room temperature. Afterthe reaction was completed, it was diluted with aqueous solution (100mL). The aqueous phase was extracted with dichloromethane (3×150 mL).The organic phases were combined, and washed with saturated salinesolution (100 mL), dried over anhydrous sodium sulfate, filtered, andconcentrated. Example 14C (55 g, 100% yield) was obtained withoutpurification, and can be directly used in the next step. ¹H-NMR (CDCl₃)δ 0.66 (3H, s, CH₃-18); 0.77 (3H, s, CH₃-26); 1.00 (3H, d, CH₃-21); 1.20(3H, s, CH₃-19); 1.96 (3H, s, AcO), 2.18-2.31 (1H, m, CH-22); 3.70 (1H,m, CH-7); 4.55 (1H, m, CH-3); 6.11 (1H, dd, =6.2 Hz, J2=8.3 Hz; CH-23);7.14-7.36 (10H, m, Ph).

Example 14D

Sodium periodate (13.2 g, 61.9 mmol) was dissolved in water (13 mL) and2N aqueous sulfuric acid solution (1.7 mL). After being stirred for 15mins, the reaction solution was cooled to 0° C., and rutheniumtrichloride (71.3 mg, 0.4 mmol) was added thereto. The reaction solutionwas stirred continusously until the color thereof turned bright yellow.Ethyl acetate (27 mL) and acetonitrile (20 mL) were added to thereaction solution, and they were stirred for another 5 min. Example 14C(4 g, 6.9 mmol) was added to the above reaction solution at 0° C., andit was filtered after the reaction was completed. The filtrate waspoured into water and extracted with ethyl acetate (3×100 mL). Theorganic phases were combined and washed with saturated sodiumthiosulfate solution (200 mL), dried over anhydrous sodium sulfate andfiltered. The filtrate was evaporated to dryness, and then isolated bysilica gel column chromatography, so as to give Example 14D (2.7 g, 89%yield). ¹H-NMR (CDCl₃) δ 0.71 (3H, s, CH₃-18); 0.86-1.07 (9H, m, CH₃-19,CH₃-21, C-24); 2.03 (3H, s, AcO); 4.48-4.61 (1H, m, CH-3).

Example 14E

Triethylamine (6.7 mL, 3.4 mmol) was added to a solution of Example 14D(1 g, 2.2 mmol) and isobutyl chloroformate (3.5 mL, 2.7 mmol) intetrahydrofuran (20 mL), and the reaction was completed after 1 hour.The reaction solution was filtered, and sodium borohydride (847 mg, 22.4mmol) was added in portions to the filtrate at 0° C. After the reactionwas completed, water (3 mL) was added to quench the reaction, and theywere stirred at room temperature for another 2 hours, followed byacidified with 3N dilute aqueous hydrochloric acid, and extracted withethyl acetate (3×15 mL). The organic phases were combined and washedwith saturated saline solution (15 mL), dried over anhydrous sodiumsulfate, and filtered. After the filtrate was evaporated to dryness,Example 14E (800 mg, 85% yield) was obtained by separation via silicagel column chromatography. ¹H-NMR (CDCl₃) δ 0.67 (3H, s, CH₃-18);0.86-0.97 (9H, m, CH₃-19, CH₃-21, CH₃-24); 2.03 (3H, s, AcO); 3.72 (3H,m, (2H, m, CH-7, CH-23); 4.48-4.61 (1H, m, CH-3).

Example 41

Sodium hydrogen (18.4 mg, 460.1 μmol, 60%) was added to a solution ofExample 14E (100 mg, 230.1 μmol) in N,N-dimethylformamide (1 mL) at 0°C. After half an hour, a solution of methyl 4-chloropicolinate (78.9 mg,460.1 μmol) in N,N-dimethylformamide (2 mL) was slowly added dropwise at0° C. After the dropwise addition was completed, the reaction system wasslowly warmed up to room temperature and reacted for 12 hours. Water (10mL) was added, and the reaction system was adjusted to pH=6 withhydrochloric acid (1 M), and extracted with dichloromethane (10 mL×3).The organic layer was dried over anhydrous sodium sulfate, filtered andevaporated to dryness. 10% sodium hydroxide solution (volume ratio ofmethanol to water was 1:1) (50 mg, 1.3 mmol, 0.5 mL) was added to asolution of the residue in methanol (1 mL), and the reaction system wasreacted at 70° C. for 2 hours. The solvent was evaporated to dryness.The system was adjusted to pH=1-2 with dilute hydrochloric acid (1 M),and extracted with dichloromethane:methanol=10:1 (10 mL×3). The organiclayer was dried over anhydrous sodium sulfate, filtered, andconcentrated. The residue was purified by preparative chromatography, soas to give Example 41 (5 mg, 4% yield). ¹H NMR (400 MHz, METHANOL-d₄) δ8.67 (br d, J=6.5 Hz, 1H), 7.99 (d, J=2.5 Hz, 1H), 7.69 (br d, J=6.5 Hz,1H), 4.50 (br d, J=6.5 Hz, 2H), 3.78-3.59 (m, 1H), 3.32-3.30 (m, 1H),2.18-1.17 (m, 25H), 1.10 (d, J=6.5 Hz, 3H), 0.95-0.89 (m, 6H), 0.76 (s,3H).

Route 15

Example 42

To a solution of Example 16D (100.0 mg, 245.9 μmol) and5-aminopyridine-3-carboxylic acid (50.0 mg, 295 μmol) in DMF (5 mL),1,3-dicyclohexylcarbodiimide (101.5 mg, 492 μmol) and4-dimethylaminopyridine (1.5 mg, 12.3 μmol) were added. The reactionsystem was reacted at 30° C. for 16 hours. A solution of lithiumhydroxide (10.3 mg, 246 μmol) in water (2.0 mL) was added to the system.The reaction system was reacted at 30° C. for 2 hours. The system wasadjusted to pH=4 with hydrochloric acid (1 M), and the aqueous phase wasextracted with dichloromethane/methanol (10:1) (20 mL×3). The organiclayers were combined, and dried over anhydrous sodium sulfate, filteredand evaporated to dryness. The residue was isolated and purified bypreparative isolation (HCl), so as to give Example 42 (20 mg, 15.4%). ¹HNMR (400 MHz, METHANOL-d₄) 9.14-9.13 (m, 1H), 8.90-8.89 (m, 1H),8.71-8.70 (m, 1H), 3.68 (s, 1H), 3.40-3.30 (m, 1H), 2.64-2.61 (m, 1H),2.15-2.06 (m, 2H), 1.99-1.20 (m, 25H), 1.07 (d, J=6.5 Hz, 3H), 0.95-0.91(m, 6H), 0.78 (s, 3H).

Route 16

Example 43

Example 16A

Reference Example 1F (10.0 g, 23.9 mmol) was dissolved intetrahydrofuran (60.0 mL). Perchloric acid (240.0 mg, 2.4 mmol, 144.6μL) (about 10 drops) was added thereto. Formic acid (40.3 g, 874.7 mmol,33.0 mL) was added dropwise within half an hour at 30° C., and thereaction solution was stirred at 50° C. for 11.5 hours. The solvent wasremoved by concentration. Water (35.0 mL) was added to the reactionsolution, which was extracted with ethyl acetate (30.0 mL×3). Theorganic layer was washed with water (10.0 mL×2), dried over anhydroussodium sulfate, filtered and concentrated, so as to give a crudeproduct. The crude product was isolated by column chromatography, so asto give Example 16A (7.0 g, 15.7 mmol, 65.6% yield, white solid). ¹H NMR(400 MHz, CHLOROFORM-d) δ=8.01 (s, 1H), 4.86-4.73 (m, 1H), 2.82-2.67 (m,1H), 2.47-2.35 (m, 2H), 2.33-2.16 (m, 2H), 2.05-1.93 (m, 2H), 1.89 (d,J=13.1 Hz, 2H), 1.82 (dd, J=5.5, 16.8 Hz, 2H), 1.75 (dd, J=6.5, 14.1 Hz,3H), 1.71 (br. s., 1H), 1.58-1.30 (m, 7H), 1.26 (s, 3H), 1.23-1.02 (m,4H), 0.95 (d, J=6.5 Hz, 3H), 0.83 (t, J=7.4 Hz, 3H), 0.68 (s, 3H).

Example 16B

Example 16A (5.8 g, 13.0 mmol) was dissolved in trifluoroacetic acid(40.0 mL) and trifluoroacetic anhydride (20.5 g, 97.4 mmol) at 0° C.After the solid was dissolved, sodium nitrite (2.7 g, 39.0 mmol) wasadded in portions, which was stirred at 0° C. for another 1 hour, andwarmed up to 40° C. and stirred for another 1 h. The reaction solutionwas cooled to 30° C., and neutralized with 0.5 mol of aqueous sodiumhydroxide solution (pH=7-8) at 0° C. The reaction solution was extractedwith ethyl acetate (40 mL×3), and the organic layer was washed withwater (10 mL), dried over anhydrous sodium sulfate, filtered andconcentrated. The crude product was isolated by chromatographic column(silica gel), so as to give Example 16B (3.5 g, 8.5 mmol, 93.0% yield,pale yellow oil). ¹H NMR (400 MHz, CHLOROFORM-d) δ=3.59-3.47 (m, 1H),2.69 (q, J=6.2 Hz, 1H), 2.42-2.31 (m, 2H), 2.29-2.15 (m, 2H), 2.01-1.88(m, 2H), 1.86-1.68 (m, 7H), 1.61-1.45 (m, 6H), 1.26 (t, J=7.2 Hz, 5H),1.19-1.12 (m, 5H), 1.02-0.91 (m, 1H), 0.80 (t, J=7.4 Hz, 3H), 0.71-0.64(m, 3H).

Example 16C

Example 16B (3.5 g, 8.5 mmol) was dissolved in methanol (100.0 mL).Aqueous potassium hydroxide solution (70.0 g, 1.3 mol, dissolved in100.0 mL of water) was added thereto, and the reaction solution wasstirred at 100° C. for 12 hours. A part of the solvent was removed byconcentration, and extracted with dichloromethane (30 mL×3). The aqueousphase was acidified with 1 mol of hydrochloric acid (pH=3-4), andextracted with ethyl acetate (30 mL×3). The organic layer was washedwith water (20 mL), dried over anhydrous sodium sulfate, filtered andconcentrated, so as to give a crude product. Example 16C (3.2 g, 7.9mmol, 93.5% yield, yellow oil) was obtained without purification. ¹H NMR(400 MHz, CHLOROFORM-d) δ=3.65-3.50 (m, 1H), 2.71 (d, J=5.8 Hz, 1H),2.48 (dd, J=2.6, 14.9 Hz, 1H), 2.43-2.31 (m, 2H), 2.22-2.15 (m, 1H),2.07-1.98 (m, 2H), 1.95-1.86 (m, 3H), 1.82-1.70 (m, 6H), 1.53-1.46 (m,3H), 1.19-1.10 (m, 6H), 1.02 (d, J=6.3 Hz, 3H), 0.86 (d, J=10.3 Hz, 5H),0.69 (s, 3H).

Example 16D

To a solution of aqueous sodium hydroxide (949.2 mg, 23.7 mmol,dissolved in 10.00 mL of water), Example 16C (3.2 g, 7.9 mmol) wasadded. The reaction solution was warmed up to 80° C., and sodiumborohydride (1.8 g, 47.5 mmol) was added in portions. The reactionsolution was stirred at 100° C. for 12 hours. Methanol (6 mL) was addeddropwise, and concentrated to remove a part of the solvent. The reactionsolution was acidified with 1 mol of hydrochloric acid (pH=5-6),extracted with ethyl acetate (40 mL×3). The organic layer was washedwith water (20 mL), dried over anhydrous sodium sulfate, filtered andconcentrated. Example 16D (3.1 g, 7.6 mmol, 96.4% yield, white solid)was obtained without isolating the crude product. ¹H NMR (400 MHz,CHLOROFORM-d) δ=3.70 (br. s., 1H), 3.46-3.36 (m, 1H), 2.52-2.39 (m, 1H),2.02-1.88 (m, 3H), 1.85-1.77 (m, 4H), 1.71-1.59 (m, 3H), 1.53-1.44 (m,4H), 1.41-1.37 (m, 1H), 1.36-1.27 (m, 4H), 1.24-1.13 (m, 4H), 1.04 (d,J=6.5 Hz, 3H), 0.92-0.88 (m, 6H), 0.73-0.69 (m, 3H).

Example 16E

To a solution of Example 16D (100 mg, 245.8 μmol) in acetonitrile (2mL), triethylamine (49.8 mg, 491.9 mmol, 68.2 μL) andN,O-dimethylhydroxylamine hydrochloride (24.0 mg, 245.9 μmol) were addedunder nitrogen atmosphere. After being stirred at 25° C. for 30 min,O-benzotriazol-N,N,N′,N′-tetramethyluronium tetrafluoroborate (98.7 mg,307.4 μmol) was then added and stirred for another 11.5 hours. Thereaction solution was poured into cold water (30 mL), and extracted withethyl acetate (40 mL×3). The organic layer was washed with water (20mL), dried over anhydrous sodium sulfate, filtered and concentrated.Example 16E (90 mg, 91.4% yield, white solid) was obtained withoutisolating the crude product. ¹H NMR (400 MHz, CHLOROFORM-d) δ=3.76 (s,3H), 3.70 (br. s., 1H), 3.46-3.36 (m, 1H), 2.79 (s, 3H), 2.43-2.39 (m,1H), 2.24-2.18 (m, 1H), 2.00-1.97 (m, 41H), 1.03-0.88 (m, 10H), 0.73 (s,3H).

Example 16F

To a solution of lithium aluminum hydride (6.3 mg, 168.9 μmol) intetrahydrofuran (2 mL), Example 16E (50 mg, 111.2 μmol) was added at−78° C., and stirred at −78° C. for 2 hours. After the reaction wascompleted, water (0.006 mL) was added and the reaction solution wasfiltered. The filter cake was oven-dried to give Example 16F (50 mg,100%, white solid). ¹H NMR (400 MHz, CHLOROFORM-d) δ=9.76 (s, 1H), 3.70(br. s., 1H), 3.46-3.36 (m, 1H), 2.43-2.39 (m, 1H), 2.24-2.18 (m, 1H),2.00-1.97 (m, 41H), 1.03-0.88 (m, 1H), 0.73 (s, 3H).

Example 16G

To a solution of Example 16F (150.0 mg, 0.3 mmol) and methyl5-aminopyridine-3-carboxylate (64.0 mg, 0.4 mmol) in ethyl acetate (5mL), trifluoroacetic acid (57 μL, 0.8 mmol) and sodiumtriacetoxyborohydride (146.0 mg, 0.7 mmol) were added. The reactionsystem was reacted at 20° C. for 16 hours. Ethyl acetate (30 mL) wasadded to the reaction system. The system was washed with saturatedsodium bicarbonate solution (20 mL×2) and saturated saline solution (20mL×1). The organic layer was dried over anhydrous sodium sulfate,filtered and evaporated to dryness, so as to give the crude product. Thecrude product was isolated by preparative isolation (TFA) to giveExample 16G (60.0 mg, 29.7%). ¹H NMR (400 MHz, CHLOROFORM-d) 8.44-8.43(m, 1H), 8.43-8.42 (m, 1H), 7.92-7.91 (m, 1H), 4.01 (s, 3H), 3.72-3.71(m, 1H), 3.49-3.48 (m, 1H), 3.28-3.13 (m, 2H), 1.99-1.20 (m, 25H), 1.01(d, J=6.5 Hz, 3H), 0.95-0.87 (m, 6H), 0.67 (s, 3H).

Example 43

To a solution of Example 16G (60.0 mg, 113.9 μmol) in tetrahydrofuran (2mL), methanol (1 mL) and water (2 mL), lithium hydroxide (60.0 mg, 1.43mmol) was added. The reaction system was reacted at 40° C. for 1 hour.The system was adjusted to pH=4 with hydrochloric acid (1 M), and theaqueous phase was extracted with dichloromethane/methanol (10:1) (20mL×3). The organic layers were combined, dried over anhydrous sodiumsulfate, filtered and evaporated to dryness. The residue was isolatedand purified by preparative isolation (HCl) to give Example 43 (40 mg,68%). ¹H NMR (400 MHz, METHANOL-d₄) 8.30-8.29 (m, 1H), 8.03-8.02 (m,1H), 7.59-7.58 (m, 1H), 3.70-3.65 (m, 1H), 3.40-3.30 (m, 1H), 3.12-3.07(m, 2H), 1.99-1.20 (m, 25H), 1.05 (d, J=6.5 Hz, 3H), 0.95-0.87 (m, 6H),0.72 (s, 3H).

Route 17

Example 44

Concentrated sulfuric acid (368.0 mg, 3.8 mmol) was added to a solutionof Example 22 (50.0 mg, 97.3 μmol) in methanol (5 mL). The reactionsystem was reacted at 70° C. for 12 hours. Water (5 mL) was added to thesystem. The system was adjusted to pH=7 with sodium hydroxide (1M), andextracted with dichloromethane (10 mL×3). The organic layer was driedover anhydrous sodium sulfate, filtered and concentrated. The residuewas purified by preparative chromatography, so as to give the titlecompound 44 (15 mg, 29.2% yield). ¹H NMR (400 MHz, METHANOL-d₄) δ 8.76(d, J=2.0 Hz, 1H), 8.18 (dd, J=2.5, 8.8 Hz, 1H), 6.82 (d, J=8.8 Hz, 1H),4.49-4.28 (m, 2H), 3.89 (s, 3H), 3.65 (br s, 1H), 3.28 (br s, 1H),2.03-1.15 (m, 25H), 1.04 (d, J=6.5 Hz, 3H), 0.93-0.86 (m, 6H), 0.71 (s,3H).

Route 18

Example 45

Example 18A

Example 18A was synthesized by using Example 23 as a raw material, andthe procedure was referred to the synthesis of Example 12A. The yieldwas 79.7%. ¹H NMR (400 MHz, CHLOROFORM-d) δ 8.48 (d, J=2.3 Hz, 1H), 8.08(d, J=2.3 Hz, 1H), 7.26 (s, 1H), 6.55 (t, J=4.8 Hz, 1H), 4.57-4.38 (m,2H), 4.24 (d, J=5.0 Hz, 2H), 3.81 (s, 3H), 3.71 (s, 1H), 3.46-3.36 (m,1H), 2.05-1.12 (m, 33H), 1.06-0.84 (m, 14H), 0.68 (s, 3H).

Example 45

The procedure of Example 45 was referred to the synthesis of Example 39.The yield was 56.8%. ¹H NMR (400 MHz, METHANOL-d₄) δ 8.80-8.72 (m, 1H),8.48 (d, J=2.0 Hz, 1H), 8.10 (d, J=2.0 Hz, 1H), 4.50-4.33 (m, 2H), 3.99(d, 1=4.5 Hz, 2H), 3.56 (s, 1H), 1.98-1.03 (m, 31H), 1.01-0.87 (m, 5H),0.84-0.74 (m, 10H), 0.62 (s, 3H).

Example 1: In Vitro Evaluation FXR Biochemical Experiment ExperimentalPurpose:

The activation effect of the compound on FXR binding reaction wasdetected by AlphaScreen.

Experimental Materials:

1. Protein: Glutathione-S-transferase-labeled FXR human protein(Invitrogen)2. Co-activator: Biotin-labeled steroid receptor coactivator (Anaspec)3. Detection reagent: AlphaScreen Detection Kit (PerkinElmer)

Experimental Method:

1. Compound Dilution: The compound to be tested was prepared as a 40 μMDMSO solution, and then diluted 3-fold to 10 concentration points. Thereference compound was prepared as a 400 μM DMSO solution, and thendiluted 1.5-fold to 10 concentration points. The diluted DMSO solutionwas added to the wells of a 384-well plate in a volume of 150 nL perwell.2. The glutathione-S-transferase-labeled FXR human protein and thebiotin-labeled steroid receptor coactivator were formulated as a mixedsolution with concentrations of 0.4 nM and 30 nM, respectively, added tothe wells of the 384-well plate in a volume of 15 μL per well, andincubated for 1 hour at room temperature.4. The mixed solution of acceptor beads in the AlphaScreen Detection Kitwas diluted 125-fold, and added to the wells of the 384-well plate in avolume of 7.5 μL per well. The operation during the experimental processwas protected from light. The incubation was performed for 1 hour atroom temperature.5. The mixed solution of donor beads in the AlphaScreen Detection Kitwas diluted 125-fold, and added to the wells of the 384 well-plate in avolume of 7.5 μL per well. The operation during the experimental processwas protected from light. The incubation was performed for 1 hour atroom temperature.6. EC50 test: Envision was used with excitation at 680 nm to read theabsorbance signals at 520-620 nm.7. Analytical data: The data were analyzed via using Prism 5.0, and theEC50 values of the activation effects of the compounds were calculated.The ratio of the highest signal value of the compound to that of thereference compound was then used to give the percentage of activationefficacy of the compound.

FXR Cell Experiment Experimental Purpose:

The effect of the compound on the cellular functional activity wasdetected by p-lactamase reporter gene technique.

Experimental Materials:

1. Cell line: FXR HEK 293T DA2. Cell culture medium: DMEM medium supplemented with 10% serum andPenicillin/Streptomycin (lx)3. Detection reagent: GeneBLAzer® Reporter Gene Detection Kit(Invitrogen)

Experimental Method:

1. Compound Dilution: The compound to be tested was prepared as a 100 μMDMSO solution, and then the compound was diluted 3-fold to 10concentration points. The reference compound was prepared as a 100 μMDMSO solution, and then diluted 1.3-fold to 10 concentration points. Thediluted DMSO solution was added to the wells of a 384-well plate in avolume of 200 nL per well. 2. Cell inoculation: FXR HEK 293T DA cellswere resuscitated, resuspended in a culture medium, diluted to a densityof 5×10⁵ cells/mL, and added to the wells of the 384-well plate in avolume of 40 μL per well.3. The 384-well microplate was incubated at 37° C., 5% CO₂ for 16 hours.4. 6 μL of 1 mM LiveBLAzer™-FRET B/G (CCF4-AM) substrate was mixed with60 μL of B solution and 934 μL of C solution, and added to the wells ofthe 384-well plate in a volume of 8 μL per well.5. The 384-well microplate was incubated in dark for 2 hours at roomtemperature.6. EC50 test: Envision was used with excitation at 409 nm to read theabsorbance signals at 460 and 530 nm.7. Analytical data: The data was analyzed via using Prism 5.0, and theEC50 values of the activation effects of the compounds were calculated.The ratio of the highest signal value of the test compound to that ofthe reference compound (chenodeoxycholic acid, CDCA) was then used togive the percentage of activation efficacy of the compound.

TABLE 1 Test results of EC₅₀ for the biochemical experiment and cellexperiment: Test Samples FXR enzyme activity FXR cell activity (Titlecompound) EC₅₀ (μM ) Efficacy EC₅₀ (μM ) Efficacy Chenodeoxycholic 12.14100% 10.22 100% acid,CDCA Example 1  0.9 320% 17.9 134% Example 2  1.46114% Example 3  2.62 106% Example 4  0.72 334% 1.87 131% Example 5  0.09134% 0.57 190% Example 6  0.36 375% 5.16 132% Example 7  0.03 360% 0.06140% Example 8  0.11 264% Example 9  0.16 407% 0.14 130% Example 10 0.06188% 0.2 138% Example 11 0.03 241% 0.1 132% Example 12 0.07 240% Example13 0.39 270% Example 14 0.73 195% Example 16 0.31 247% Example 17 0.60313% 2.50 131% Example 18 0.07 187% Example 19 0.15 251% 0.37 143%Example 20 0.06 287% Example 22 0.006 249% Example 23 0.0025 248% 0.003150% Example 24 0.0025 138% Example 25 0.011 233% Example 26 0.13 280%Example 27 0.006 212% Example 28 0.007 219% Example 29 0.012 190%Example 30 0.056 150% Example 31 0.027 204% Example 32 0.650 140%Example 33 0.141 194% Example 34 0.093 191% Example 38 0.02 210% Example39 0.02 205% Example 40 0.045 197% Example 41 0.08 127% Example 42 1.43121% Example 43 0.37 193% Example 44 0.330 140%

Conclusion: The agonistic effect of the compound of the presentinvention on FXR receptor is significant, and the agonistic effect onFXR receptor at the cellular level is also significant.

Experimental Example 2: In Vivo Study

Pharmacokinetics in mice administrated with single compound:

12 male mice (C57BL/6J) were randomly divided into two groups, i.e., 6mice per group. The first group was the intravenous administrationgroup, involving administration at a dose of 2 mg/kg, 2 mL/kg byinjecting via tail vein (the vehicle was 10% HPbCD aqueous solution, andif the drug solubility was not satisfactory, the cosolvent was added);the second group was the oral administration group, involvingintragastrical administration at a dose of 10 mg/kg, 10 mL/kg (thevehicle was 0.5% HPMC aqueous solution). Plasma (using K₂-EDTA asanticoagulant) samples were taken at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8 and24 hours in the intravenous administration group after administration;and plasma samples were taken at 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hoursin the oral administration group after administration. For 6 animals ineach group, blood samples were collected for 3 animals at one timepoint. The first batch of 3 animals was alternately sampled with thesecond batch of 3 animals. Plasma sample analysis was performed by usingLC-MS/MS. The resultant plasma concentrations were plotted with repeatto time, and PK parameters were calculated by using Phoenix WinNonlin6.3.

TABLE 2 Ex- Ex- Ex- Compound INT-747 ample 11 ample 22 ample 23 Dosage(mg/kg) 10 10 10 10 PK Cmax (nM) 1013 1433 936 1777 parameters Tmax (h)0.3 2 0.5 0.5 in plasma AUC 993 5173 2337 1109 (nM · h) F% 13% — 34% 20%

Conclusion: As shown in Table 2, after oral administration at the samedosage, the peak concentration and the drug exposure of compound 11 werehigher than those of the control compound INT-747. After oraladministration at the same dosage, the peak concentration of compound 22was close to that of the control compound INT-747, and the drug exposureof compound 22 was higher than that of the control compound INT-747.After oral administration at the same dosage, the peak concentration ofcompound 23 was higher than that of the control compound INT-747, andthe drug exposure of compound 23 was also higher than that of thecontrol compound INT-747.

Liver-Blood Ratio Experiment of Mice Via Cassette Dosing

6 male mice (C57BL/6J) were grouped as an oral administration group. 5kinds of the developed drugs were contained in the formulation, andintragastrical administration was performed at a dose of 2mg/kg/compound (the vehicle was 0.5% HPMC aqueous solution). The fivecompounds were firstly and respectively dissolved in the vehicle, andsonicated or whirled to form a 1 mg/mL solution (clear solution orsuspension), respectively; and then the solutions of the five compoundwere mixed in equal volumes (1:1:1:1:1, v:v:v:v:v) into a glass bottle.After intragastrical oral administration, plasma and liver tissuesamples were collected from 3 animals at 0.5 h after administration;corresponding samples were collected from the other 3 animals at 3 hafter administration. After collection, the liver tissue was homogenizedby using ice-cold homogenization buffer (methanol:15 mM PBS buffer (pH7.4)=1:2, v:v) based on the ratio of liver weight:homogenization buffervolume=1:3. Plasma and liver tissue samples were analyzed by using afive-in-one LC-MS/MS analysis method developed in advance. Theconcentrations of plasma and liver tissue homogenate were obtained, andthe concentration ratios of liver tissue to plasma were calculated byusing Excel.

TABLE 3 Example Example Compound INT-747 22 26 Dosage (mg/kg) 2 2 2 PKConcentration in liver ( nM ) 0.5 h / 3 h 711/625 1959/701 3904/358parameters Concentration in plasma ( nM ) 0.5 h / 3 h 151/63   83/45  387/18  Concentration ratio of liver to plasma 0.5 h / 3 h  5/10  24/16   10/20  Note: ND indicates “not detected”.

Conclusion: As shown in Table 3, after oral administration of thecompound in the present invention at the same dosage, the drugconcentrations of Example 22 in the liver at 0.5 h and 3 h were higherthan those of the control compound, and the liver/blood concentrationratios were also higher than those of the control compound at 0.5 h and3 h. The drug concentration of Example 26 in the liver at 0.5 h washigher than that of the control compound, and the liver/bloodconcentration ratios of Example 26 were higher than those of the controlcompound at 0.5 h and 3 h.

1. A compound represented by formula (I),

wherein, ring A is selected from 5- to 12-membered aryl, 5- to12-membered heteroaryl containing 1 to 2 heteroatoms, 5- to 6-memberednon-aromatic heterocyclyl containing 1 to 2 heteroatoms or 5- to6-membered cycloalkyl, and said ring A is optionally substituted with 1,2 or 3 R_(a), or with 1, 2 or 3 oxo groups, and said heteroatom isselected from N, O or S; L is selected from C₁₋₈ alkyl, C₁₋₈ heteroalkylor C₂₋₈ alkenyl, and said L is optionally substituted with 1, 2 or 3R_(b), or with 1, 2 or 3 oxo groups; L₁ is selected from O, N(R_(d)),N(R_(d))S(═O)₂ or N(R_(d))S(═O); R₁ is selected from H, C₁₋₆ alkyl, C₁₋₆heteroalkyl, 5- to 6-membered aryl, 4- to 6-membered heteroaryl, C₃₋₆cycloalkyl, or 3- to 6-membered heterocycloalkyl, and said C₁₋₆ alkyl,C₁₋₆ heteroalkyl, 5- to 6-membered aryl, 4- to 6-membered heteroaryl,C₃₋₆ cycloalkyl, or 3- to 6-membered heterocycloalkyl is optionallysubstituted with 1, 2 or 3 R_(c), or with 1, 2 or 3 oxo groups; R_(a),R_(b), R_(c) and R_(d) are each independently selected from H, F, Cl,Br, I, OH, CN, NO₂, NH₂, COOH, S(═O)₂OH, C₁₋₃ alkylamino, N,N-di(C₁₋₃alkyl)amino, C₁₋₃ alkyl, C₁₋₃ alkyloxy or C₁₋₃ alkylthio, and said C₁₋₃alkylamino, N,N-di(C₁₋₃ alkyl)amino, C₁₋₃ alkyl, C₁₋₃ alkyloxy or C₁₋₃alkylthio is optionally substituted with 1, 2 or 3 R′; R′ is selectedfrom F, Cl, Br, I, OH, NH₂, NO₂, CN, COOH, Me, Et, CH₂F, CHF₂, CF₃,CH₃O, CH₃S, NH(CH₃) or N(CH₃)₂; or a pharmaceutically acceptable saltthereof.
 2. The compound or the pharmaceutically acceptable salt thereofaccording to claim 1, wherein R_(a), R_(b), R_(c) and R_(d) are eachindependently selected from H, F, Cl, Br, I, COOH, S(═O)₂OH, Me, CF₃,CHF₂, CH₂F, Et, OMe, NH(CH₃) or N(CH₃)₂.
 3. The compound or thepharmaceutically acceptable salt thereof according to claim 1, whereinring A is selected from phenyl, pyridyl, pyridin-2(1H)-onyl, pyrimidyl,pyrazolyl, imidazolyl, oxazolyl, thiazolyl, thienyl, pyrrolidinyl,piperidinyl, morpholinyl, piperazinyl, isoxazolyl, isothiazolyl,bicyclo[1.1.1]pentyl, benzoxazolyl, benzo[d]isoxazolyl, indazolyl,indolyl, quinolinyl, isoquinolinyl, quinazolinyl,1H-pyrrolo[2,3-B]pyridyl, indolizinyl, benzothiazolyl or benzothienyl,and said ring A is optionally substituted with 1, 2 or 3 R_(a).
 4. Thecompound or the pharmaceutically acceptable salt thereof according toclaim 3, wherein ring A is selected from

and said ring A is optionally substituted with 1, 2 or 3 R_(a).
 5. Thecompound or the pharmaceutically acceptable salt thereof according toclaim 4, wherein ring A is selected from


6. The compound or the pharmaceutically acceptable salt thereofaccording to claim 1, wherein L is selected from C₁₋₄ alkyl, C₁₋₄alkoxy, C₁₋₄ alkylthio, C₁₋₄ alkylamino, C₁₋₄ alkyl-C(═O)NH—, C₂₋₄alkenyl or C₁₋₃ alkyl-O—C₁₋₃ alkyl, and said L is optionally substitutedwith 1, 2 or 3 R_(b).
 7. The compound or the pharmaceutically acceptablesalt thereof according to claim 6, wherein L is selected from

and said L is optionally substituted with 1, 2 or 3 R_(b).
 8. Thecompound or the pharmaceutically acceptable salt thereof according toclaim 7, wherein L is selected from


9. The compound or the pharmaceutically acceptable salt thereofaccording to claim 8, wherein L is selected from


10. The compound or the pharmaceutically acceptable salt thereofaccording to claim 1, wherein L₁ is selected from O, NH, NHS(═O)₂ andNHS(═O).
 11. The compound or the pharmaceutically acceptable saltthereof according to claim 1, wherein R₁ is selected from H, C₁₋₃ alkyl,C₃₋₆ cycloalkyl, phenyl, pyridyl, pyridazinyl, pyrazinyl, imidazolyl,pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl or thienyl, andsaid C₁₋₃ alkyl, C₃₋₆ cycloalkyl, phenyl, pyridyl, pyridazinyl,pyrazinyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl,isoxazolyl or thienyl is optionally substituted with 1, 2 or 3 R_(c),and said R_(c) is as above defined.
 12. The compound or thepharmaceutically acceptable salt thereof according to claim 11, whereinR₁ is selected from H, Me,


13. The compound according to claim 1, wherein it is selected from:


14. A pharmaceutical composition comprising a therapeutically effectiveamount of the compound or the pharmaceutically acceptable salt thereofaccording to claim 1, and a pharmaceutically acceptable carrier.
 15. Amethod of treating Farnesoid X Receptor related diseases, cholestaticliver diseases, fibrotic diseases, hypercholesterol diseases,hypertriglyceride diseases and cardiovascular diseases, comprisingadministering a therapeutically effective amount of the compound or thepharmaceutically acceptable salt thereof according to claim
 1. 16. Themethod according to claim 15, wherein the diseases are selected fromnon-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis(NASH), primary biliary cirrhosis (PBC), cholestatic hepatopathy,chronic liver disease, hepatitis C infection, alcoholic liver disease,hepatic fibrosis, primary sclerosing cholangitis (PSC), gallstone,biliary atresia, lower urinary tract symptom and benign prostatichyperplasia (BPH), ureteral calculi, obesity, type 2 diabetes,atherosclerosis, atherosclerotic symptom, hepatic function injuryresulting from hypercholesterolemia and hyperlipidemia.